• Asian Pacific Journal of Cancer Prevention
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• 제17권1호
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• pp.109-115
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• 2016

Background: P53 has been reported to be involved with tumorigenesis and has also been implicated as a significant biomarker in oral squamous cell carcinoma(OSCC). However, the diagnostic value of p53 antibodies remains controversial; hence, we comprehensively and quantitatively assessed the potential in the present systematic review. Materials and Methods: A comprehensive search was performed using PubMed and Embase, up to October 31, 2014, without language restriction. Studies were assessed for quality using QUADAS (quality assessment of studies of diagnostic accuracy). The positive likelihood ratio (PLR) and negative likelihood ratio (NLR) were pooled separately and compared with overall accuracy measures using diagnostic odds ratios (DORs) and symmetric summary receiver operating characteristic (SROC) curves. Results: Of 150 studies initially identified, 7 eligible regarding serum p53 antibodies met the inclusion criteria. Some 85.7% (6/7) were of relatively high quality (QUADAS $score{\geq}7$). The summary estimates for quantitative analysis of serum p53 antibody in the diagnosis of squamous cell carcinoma were: PLR 2.06 [95% confidence interval (CI) : 1.35-3.15], NLR 0.85 (95%CI: 0.80-0.90) and DOR 2.47 (95%CI: 1.49-4.12). Conclusions: This meta-analysis suggests that the use of s-p53-antibodies has potential diagnostic value with relatively high sensitivity and specificity for OSCC particularly with serum specimens for discrimination of OSCCs from healthy controls. However, its discrimination power is not perfect because of low sensitivity.

• 대한세포병리학회지
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• 제7권2호
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• pp.138-143
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• 1996

The diagnostic accuracy of routine cytological preparations from effusions ranges from 60% to 70%. Immunohistochemical markers, especially tumor-associated antigens, have been successfully employed to increase diagnostic sensitivity in effusion cytology. However, more than two different antibodies in diagnosis of effusions are needed. In the view of prevalence of abnormalities of p53 gene in human malignancies we investigated the diagnostic usefulness of demonstration of p53 protein immunoreactivity in distinguishing benign changes versus malignant processes in effusions. p53 protein expression was studied immunohistochemically in 76 effusions(28 malignant and 48 benign) using anti-human p53 antibody p53 immunoreactivity was identified in 19 of 28(67.9%) malignant effusions. In contrast, no p53 immunoreactivity was observed in all benign effusions. A specificity of 100% and a sensitivity of 67.9% were observed. These results suggest that immunohistochemical detection of p53 protein seems to be helpful in distinguishing benign changes versus malignant processes in effusions, although its principal limitation is its relatively low sensitivity.

• Journal of Applied Biological Chemistry
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• 제53권2호
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• pp.112-115
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• 2010

To analyze the surface properties of bacteriophytochrome (BphP), five (2B8, 2C11, 3B2, 3D2, 3H7) anti-BphP monoclonal antibodies were produced by using full-length of BphP of Deinococcus radiodurnas. 2B8 and 2C11 preferentially recognized the epitopes at N-terminal region of BphP, whereas 3B2, 3D2 and 3H7 showed preferential affinities to the epitopes of C-terminal region of BphP.

• Parasites, Hosts and Diseases
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• 제39권1호
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• pp.67-76
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• 2001

This experiment was focused on the characterization of anti- Toxoplasma monoclonal antibodies (mAbs) and the effect of mAbs on the parasite invasion of mouse peritoneal macrophages. Twenty eight mAbs including M110, M556, R7A6 and M62l were characterized by Ab titer, immunoglobulin isotyping and western blot pattern. Antibody titer (optical density) of 4 mAbs. Ml 10. M556. R7A6 and M62l. were 0.53,0.67, 0.45 and 0.39 (normal mouse serum; 0.19) with the same IgGl isotypes shown by Enzyme-linked immunosorbent assay (ELISA). Western blot analysis showed that Ml 10. M556. R7A6 and M62l reacted with the 33 kDa (p30),31 kDa (p28),43 kDa and 36 kDa protein. Immuno-gold labelling of mAbs M110, M556, R7A6 and M621 reacted with the surface membrane, dense granules and parasitophorous vacuolar membrane (PVM) , rhoptries and cytoplasm of tachyzoite, respectively. For in vitro assay, preincubation of tachyzoties with four mAbs, Ml 10, M556, R7A6 and M62l resulted in the decrease of the number of infected macrophages (p < 0.05) and the suppression of parasite multiplication at 18 h post-infection. Four monoclonal antibodies including Ml 10 (SAGI) were found to have an important role in the inhibition of macrophage invasion and T. gondii multiplication in vitro, and these mAbs may be suitable for vaccine candidates, diagnostic kit and for chemotherapy.

• 미생물학회지
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• 제31권4호
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• pp.279-285
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• 1993

p53 유전자의 변화는 인간의 여러 암에서 가장 흔하게 발견되며 종양세포내에서는 이러한 변형된 p53 단백질의 양의 증가가 초래된다. 세포내에 축적된 p53 단백질의 발견은 인간의 암증세를 판단할 유용한 기중이 되기도 한다. 본 연구에서는 이러한 면역조직화학 검사에 쓰일 수 있는 폴리클로날 항체를 만들기 위햐여 사람의 p53 유전자를 glutathione S-transferase 와의 융합 단백질의 형태로서 대장균내에서 발현시켰다. p53 의 아미노산 1-158번을 코딩하고 있는 NeoI fragment 와 아미노산 159-393 번을 코딩하는 NocI-BamHI fragment 를 BamHI linker 를 이용하여 in frame 으로 pGEX-2T 의 BamHI 자리에 삽입하여 재조합 플라스미드 pGTNS 와 pGTNL 을 각각 만들었다. 또 PCR 에 의한 증폭에 의햐여 아미노산 38-145번을 코딩하는 유전자 부위를 증폭하였으며 BamHI 과 PvuII 로 절단하여 pGEX-2T의 BamHI 과 SmaI 자리에 삽입함으로써 pGTBP 를 제조하였다. 이들 재조합 균주들을 IPTG 로 4시간 induction 한 후 세포 추출물로부터 glutathione Sepharose bead 를 이용하여 융합단백질을 분리하였다. Bead 에 결합된 단백질은 10% SDS-polyacrylamide gel 에서 전기영동하였으며, 각각의 분자량은 54 kDa, 53 kDa 와 40 kDa 였다. 이러한 방법으로 1리터 배양으로부터 약 1mg 의 단백질을 정제하였다.

• Journal of the Korean Wood Science and Technology
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• 제21권3호
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• pp.87-93
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• 1993

This study was undertaken to investigate the characteristics and the productivities of lignin olytic enzymes: laccase (Lac) and Mn-dependent peroxidase (MnP) from Coriolus versicolor and Lentinus edodes respectively. Enzymes were isolated from cultural filterates and purified according to the standard methods. These enzymes showed one band in SDS-PAGE and their molecular weights were found 62,000 and 45,000 dalton respectively. Polyclonal antibodies against Lac and MnP were raised against mouse. In the ELISA (enzyme-linked immunosorbent assay), Lac and MnP-antiserum produced a strong positive reaction with Lac and MnP antigen($A_{405}$=2.50 and 3.53 respectively). The sera to negative (S/N) ratio was determined by the dividing the mean absorbance of antibodies by the corresponding diluted samples from normal mouse serum. The sera produced showed 2 times more positive reaction in S/N ratio than negative sera.

• Asian Pacific Journal of Cancer Prevention
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• 제13권4호
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• pp.1341-1347
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• 2012

Introduction: Diffuse large B-cell lymphomas (DLBCL) can be divided into germinal centre (GC-DLBCL) and post germinal centre (post GC-DLBCL) groups by applying immunohistochemical antibodies. As these subgroups respond differently to chemotherapy, it is possible at diagnosis to select a poor prognostic subgroup for aggressive treatment. Objective: To determine the frequencies of GC-DLBCL and post GC-DLBCL in patients by immunohistochemistry (IHC) and the clinical response after six cycles of chemotherapy. Subjects and Methods: In this descriptive study conducted in AFIP and CMH, Rawalpindi and NORI, Islamabad, from September 2010 to September 2011, a total of 75 pretreatment cases of DLBCL diagnosed during the study period were included. Cases were segregated in to GC-DLBCL and post GC-DLBCL groups according to results of immunohistochemistry markers CD10, BCL6 and MUM1. Immediate clinical response was assessed after 6 cycles of chemotherapy. Response was divided into complete response, partial response, stable disease or relapse or progression. Results: The mean age was $54.2{\pm}15$. Males were 53 (70.7%). Forty (53.3%) cases comprised the GC-DLBCL group; 25(62.5%) of them showed a complete response. Most patients of the post GC-DLBCL 19(54%) showed relapse/progression. Results of immediate clinical response in both prognostic subgroups were significant (p<0.05). Results regarding positivity with immunohistochemical antibodies CD10 (p 0.011), BCL6 (p 0.013) and MUM1 (p 0.000) regarding immediate clinical response were also significant. Conclusion: GC-DLBCL group shows better response to CHOP chemotherapy regimen. Immunohistochemistry should be used to further classify DLBCL as this can enable us to select aggressive group for aggressive treatment. This manuscript is important because the study is the first to becarried out exclusively in Pakistan or our part of the world.

• Parasites, Hosts and Diseases
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• 제53권1호
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• pp.29-34
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• 2015

The aim of this cross sectional case control study was to examine the serofrequency and serointensity of Toxoplasma gondii (Tg) IgG, IgM, and DNA among patients with schizophrenia. A total of 101 patients with schizophrenia and 55 healthy controls from Sungai Buloh Hospital, Selangor, Malaysia and University Malaya Medical Center (UMMC) were included in this study. The diagnosis of schizophrenia was made based on the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV). The presence of Tg infection was examined using both indirect (ELISA) and direct (quantitative real-time PCR) detection methods by measuring Tg IgG and IgM and DNA, respectively. The serofrequency of Tg IgG antibodies (51.5%, 52/101) and DNA (32.67%, 33/101) among patients with schizophrenia was significantly higher than IgG (18.2%, 10/55) and DNA (3.64%, 2/55) of the controls (IgG, P=0.000, OD=4.8, CI=2.2-10.5; DNA, P=0.000, OD=12.9, CI=2.17-10.51). However, the Tg IgM antibody between patients with schizophrenia and controls was not significant (P>0.005). There was no significant difference (P>0.005) in both serointensity of Tg IgG and DNA between patients with schizophrenia and controls. These findings have further demonstrated the strong association between the active Tg infection and schizophrenia.

• Journal of Microbiology and Biotechnology
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• 제24권12호
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• pp.1654-1663
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• 2014

To examine the effect of tumor suppressor protein p53 on the antitumor activity of 2-methoxyestradiol (2-MeO-$E_2$), 2-MeO-$E_2$-induced cell cycle changes and apoptotic events were compared between the human colon carcinoma cell lines HCT116 ($p53^{+/+}$) and HCT116 ($p53^{-/-}$). When both cell types were exposed to 2-MeO-$E_2$, a reduction in the cell viability and an enhancement in the proportions of $G_2/M$ cells and apoptotic sub-$G_1$ cells commonly occurred dose-dependently. These 2-MeO-$E_2$-induced cellular changes, except for $G_2/M$ arrest, appeared to be more apparent in the presence of p53. Immunofluorescence microscopic analysis using anti-${\alpha}$-tubulin and anti-lamin B2 antibodies revealed that after 2-MeO-$E_2$ treatment, impaired mitotic spindle network and prometaphase arrest occurred similarly in both cell types. Following 2-MeO-$E_2$ treatment, only HCT116 ($p53^{+/+}$) cells exhibited an enhancement in the levels of p53, p-p53 (Ser-15), $p21^{WAF1/CIP1}$, and Bax; however, the Bak level remained relatively constant in both cell types, and the Bcl-2 level decreased only in HCT116 ($p53^{+/+}$) cells. Additionally, mitochondrial apoptotic events, including the activation of Bak and Bax, loss of ${\Delta}{\psi}m$, activation of caspase-9 and -3, and cleavage of lamin A/C, were more dominantly induced in the presence of p53. The Bak-specific and Bax-specific siRNA approaches confirmed the necessity of both Bak and Bax activations for the 2-MeO-$E_2$-induced apoptosis in HCT116 cells. These results show that among 2-MeO-$E_2$-induced apoptotic events, including prometaphase arrest, up-regulation of Bax level, down-regulation of Bcl-2 level, activation of both Bak and Bax, and mitochondria-dependent caspase activation, the modulation of Bax and Bcl-2 levels is the target of the pro-apoptotic action of p53.

• 한국동물위생학회지
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• 제36권1호
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• pp.53-56
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• 2013

This study was carried out to investigate the seroprevalence of Neospora caninum infection in dairy cattle of northern Gyeonggi province in Korea. A total of 716 dairy cattle from 14 dairy farms were tested for the presence of antibodies against N. caninum using a commercial ELISA test kit. Herd and individual seroprevalences of N. caninum were 92.8% (13/14) and 23.1% (166/716), respectively. The seroprevalence of dairy cattle according to age was the highest at 49~72 months (27.4%), but the difference was not statistically significant (P>0.05). No significant relationship between N. caninum seropositivity with the herd size of the dairy farms was found (P>0.05). The results indicate that N. caninum infection spreads widely in dairy farms of northern Gyeonggi province in Korea.