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Robert H Getzenberg - One of the best experts on this subject based on the ideXlab platform.
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colon cancer specific Nuclear Matrix Protein alterations in human colonic adenomatous polyps
Journal of Cellular Biochemistry, 2004Co-Authors: Gisela Brunagel, Robert E Schoen, Robert H GetzenbergAbstract:Most colon cancers arise within preexisting adenomatous polyps or adenomas. The slow evolution from the non-invasive premalignant lesion, the adenomatous polyp, to invasive cancer supports a strategy of early detection. Recently, we identified unique Nuclear Matrix Proteins (NMPs) specific for colon cancer (CC2, CC3, CC4, CC5). Most of the NMPs identified are common to all cell types, but several identified NMPs are tissue and cell line specific. The objective of this study is to describe and characterize the NMP profile of premalignant adenomatous colon polyps. Specifically when in the adenoma-carcinoma sequence four specific colon cancer NMPs, previously described, appear. Using two-dimensional (2-D) gel analysis 20 colon polyps (one juvenile polyp, six tubular adenoma (TA), seven tubulovillous adenoma (TVA), six TVA with focal high-grade dysplasia (HGD), were analyzed for the presence of four (CC2, CC3, CC4, CC5) specific NMPs. CC2 was not seen in any of the premalignant polyps. CC5 was present in only two premalignant TVA with HGD and in one TA. CC3 and CC4 were present in most adenomas. None of the NMPs were seen in the juvenile polyp, which is not considered to be a precursor of colon cancer. CC2 and CC5 are NMPs expressed at the junction of an advanced adenoma and invasive colorectal cancer. CC3 and CC4 are expressed earlier in the evolution of adenomatous polyps. Development of an assay to these Proteins may serve as a new method for early detection of colorectal cancer.
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identification of calreticulin as a Nuclear Matrix Protein associated with human colon cancer
Journal of Cellular Biochemistry, 2003Co-Authors: Gisela Brunagel, Robert E Schoen, Uzma S Shah, Robert H GetzenbergAbstract:Colon cancer is one of the most common malignancies among populations in the United States and Western Europe, and one of the leading causes of worldwide morbidity and mortality due to cancer. The early detection of colon cancer is central to the effective treatment of this disease and early detection markers are needed. We have demonstrated that high-resolution two-dimensional gel analysis of Nuclear Matrix Proteins (NMPs) demonstrated a specific oncological fingerprint of colon cancer. Utilizing this approach, four Proteins specific for colon cancer was identified. Additionally, one Protein was expressed much more strongly in colon cancer compared to adjacent and normal donor tissue. The amino acid composition of this Protein revealed sequence similarity with calreticulin. The multi-functional Protein, calreticulin, is normally found in the lumen of the endoplasmic reticulum although some reports have described a Nuclear localization of the Protein. The aim of this study was to confirm the identity of the Protein as calreticulin as well as to evaluate the localization of calreticulin in the Nuclear Matrix of colon cancer tissue.
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Nuclear Matrix Protein alterations associated with colon cancer metastasis to the liver
Clinical Cancer Research, 2002Co-Authors: Gisela Brunagel, Barbara N Vietmeier, Anthony J Bauer, Robert E Schoen, Robert H GetzenbergAbstract:Purpose: The development of colon cancer markers that can detect liver metastases early and predict which patients are at risk to develop liver metastases would have a major impact on this disease. We have previously identified G. Brunagel, et al. , Cancer Research, 62: 2437–2442, 2002, Nuclear Matrix Proteins (NMPs), which are associated with colon cancer. The objective of this study is to identify the existence of a specific NMP “fingerprint” for human liver metastasis from colon cancer. Experimental Design: Using high-resolution two-dimensional gel electrophoresis, we analyzed the NMP expression of 12 matched liver metastases and adjacent normal samples and three normal donor liver samples. These were compared with colon cancer NMP patterns, along with several primary cell systems and lines. Results: Analysis of multiple gels for each sample revealed three Proteins present in all liver metastases, which are not present in normal liver tissue and normal hepatocytes. These three Proteins were also present in colon cancer samples. Conclusion: Data provided here demonstrate that the NMP composition is able to differentiate liver metastases from normal liver tissue and normal hepatocytes and that these Proteins are also expressed in colon cancer. These results further show that the adjacent normal liver tissue changes its NMP pattern of expression. Development of an assay to detect these specific NMPs in tissue and/or serum specimens is a promising modality for early detection of liver metastases from colon cancer or potentially as a prognostic tool. In addition the functional characterization of these Proteins will significantly enhance our understanding of the development of liver metastases of this disease.
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identification of Nuclear Matrix Protein alterations associated with human colon cancer
Cancer Research, 2002Co-Authors: Gisela Brunagel, Barbara N Vietmeier, Anthony J Bauer, Robert E Schoen, Robert H GetzenbergAbstract:The early diagnosis of colorectal cancer and the early detection of recurrence are central to effective treatment, as prognosis is directly related to the stage of the disease. When colorectal cancer is diagnosed at an early, localized stage, 5-year survival is 90%. With regional lymph node metastases, survival drops to 45–60%, and with distant metastases, 5-year survival is
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detection of bladder cancer using a novel Nuclear Matrix Protein blca 4
Clinical Cancer Research, 2000Co-Authors: Badrinath R Konety, Walter M Stadler, Robert H Getzenberg, Thu Suong T Nguyen, Rajiv Dhir, Roger Day, Michael J BecichAbstract:We have identified previously six Nuclear Matrix Proteins (NMPs) that are bladder cancer specific. In this study, we analyzed the expression of one of these Proteins, BLCA-4, in bladder tumors and normal bladder tissue. We also examined the appearance of BLCA-4 in the urine as a biomarker for bladder cancer. BLCA-4 was isolated from Nuclear Matrix preparations of bladder tumors, and its peptide sequence was determined. The antibodies generated against the resulting BLCA-4 peptides were then used to detect its presence in immunoblots and in urine samples by immunoassay. We analyzed tissue samples of bladder tumor and normal donor bladders and urine obtained from 51 normal individuals and 54 patients with pathologically confirmed bladder cancer. The BLCA-4 peptide sequences do not resemble any known human Protein sequences. On immunoblot analysis, BLCA-4 expression was detectable in tumor and normal tissues from patients with bladder cancer but not in any of the normal bladder tissue obtained from organ donors. Using a prospectively determined cutoff level of 13 A (absorbance) units/μg Protein, all 51 normal individuals tested were negative for BLCA-4 expression, whereas 53 of 55 samples from patients with bladder cancer were positive. These results suggest that BLCA-4 is present throughout the bladder in both the tumor and morphologically normal areas in bladder cancer patients. BLCA-4 is a very sensitive (96.4%) and specific (100%) marker for bladder cancer. BLCA-4 is a bladder cancer-specific marker that can be detected using a urine-based assay and can be used in the diagnosis of bladder cancer.
Shahrokh F Shariat - One of the best experts on this subject based on the ideXlab platform.
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assessing the clinical benefit of Nuclear Matrix Protein 22 in the surveillance of patients with nonmuscle invasive bladder cancer and negative cytology a decision curve analysis
Cancer, 2011Co-Authors: Shahrokh F Shariat, Caroline O S Savage, Thomas F Chromecki, Maxine Sun, Douglas S Scherr, Richard K Lee, Giovanni Lughezzani, Mesut RemziAbstract:BACKGROUND: Several studies have demonstrated that abnormal levels of Nuclear Matrix Protein 22 (NMP22) are associated with bladder cancer and have led to the approval of NMP22 as a urinary biomarker by the US Food and Drug Administration. Nonetheless, the clinical significance of NMP22 remains unclear. The objective of this study was to use decision analysis to determine whether NMP22 improves medical decision-making. METHODS: The current study included 2222 patients who had a history of nonmuscle-invasive bladder cancer and current negative cytology. The authors developed models to predict cancer recurrence or progression to muscle-invasive disease using voided NMP22 levels, cystoscopy, age, and sex. Clinical net benefit was calculated by summing the benefits (true-positives), subtracting the harms (false-positives), and weighting these values by the threshold probability at which a patient or clinician would opt for cytoscopy. RESULTS: After cystoscopy, 581 patients (26%) had cancer identified. The NMP22 level was associated significantly with bladder cancer recurrence and progression (P 8% for recurrence and > 3% for progression. Only offering cystoscopy to those who had a risk > 15% reduced the number of cystoscopies by 229 while missing only 25 cancer recurrences per 1000 men with negative cytology. The current study was limited by its multicenter design. CONCLUSIONS: For clinicians who would perform a cystoscopy at a threshold of 5% for recurrence or 1% for progression, NMP22 did not aid clinical decision-making. For less risk-averse clinicians who would only perform a cystoscopy at a threshold probability >thinsp;8% for recurrence or > 3% for progression, NMP22 helped to indicate which patients required cystoscopy and which could be spared this procedure. Cancer 2011. © 2011 American Cancer Society.
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impact of clinical factors including a point of care Nuclear Matrix Protein 22 assay and cytology on bladder cancer detection
BJUI, 2009Co-Authors: Yair Lotan, Umberto Capitanio, Shahrokh F Shariat, Georg C Hutterer, Pierre I KarakiewiczAbstract:OBJECTIVE To determine whether the Nuclear Matrix Protein-22 (NMP22) assay can improve the accuracy of discriminating between high-risk patients with and without bladder cancer. PATIENTS AND METHODS Age, gender, race, smoking status, haematuria and its extent, and the NMP22 and urinary cytology results, were available for 1272 patients. The data of 670 (52.7%) from four study sites were used to develop a logistic regression model-based nomogram to predict the presence of bladder cancer. The remaining data from 602 (47.3%) patients from nine study sites were used to externally validate the nomogram. A separate nomogram was developed for urinary cytology, and for the combination of NMP22 and urinary cytology findings. RESULTS Of 1272 patients, 76 (6.0%) had bladder cancer, 217 (17.1%) were NMP22-positive and 17 (1.3%) had malignant cells on urinary cytology. NMP22 and urinary cytology results were independent predictors of bladder cancer (P = 0.005 and 0.007, respectively). In external validation, the area under the curve (AUC) for NMP22 was 76.0% vs 56.2% for cytology. External validation of the multivariable NMP22-based bladder cancer nomogram gave an AUC of 82.4% vs 74.7% for the multivariable cytology-based nomogram (gain 7.7%; P = 0.006) vs 82.6% for the multivariable nomogram combining NMP22 and cytology results (gain 0.2%; P = 0.1). CONCLUSIONS The ability of the NMP22 test to predict bladder cancer in high-risk patients significantly exceeds that of urinary cytology. The NMP22-based nomogram can help to identify individuals at risk of bladder cancer.
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impact of risk factors on the performance of the Nuclear Matrix Protein 22 point of care test for bladder cancer detection
BJUI, 2008Co-Authors: Yair Lotan, Shahrokh F ShariatAbstract:OBJECTIVES: To evaluate the impact of different risk factors and symptoms on the performance characteristics of the Nuclear Matrix Protein 22 (NMP22) BladderChek test (Matritech Inc., Newton, MA, USA) for the detection of bladder cancer, as this is a point-of-care assay that measures NMPs in voided urine. PATIENTS AND METHODS: In all, 23 academic, private practice and veterans' facilities in 10 states prospectively enrolled consecutive patients from September 2001 to May 2002. Participants included 1328 patients at elevated risk of bladder cancer from factors such as a history of smoking or symptoms including haematuria and/or dysuria. Patients at risk of malignancy of the urinary tract provided a voided urine sample for analysis of NMP22 Protein and cytology before cystoscopy. RESULTS: Of the 1328 patients: no urinary disease, benign disease and malignancy were found in 545 (41%), 704 (53%) and 79 (6%) patients, respectively. Overall, the positive predictive value (PPV) for detection of bladder cancer was 20.3% (45/222) and negative predictive value (NPV) was 96.9% (1072/1106). The PPV was higher in men (24.0%) than women (13.2%). In men, the PPV increased with smoking (35.4%), gross haematuria (51.2%) and a combination of both factors (70.6%). The impact on the PPV of smoking (9.7%) and gross haematuria (28.6%) was less dramatic in women. The PPV increased from 16.8% in patients aged 65 years. The NPV remained almost always >95% except in men with gross haematuria where it decreased to 77% in smokers and 94% in non-smokers. CONCLUSION: The PPV of the BladderChek test improves in patients at higher risk of bladder cancer reaching 77% in men presenting with gross haematuria who are aged >65 years and smoke. The NPV is highest in women aged <65 years, up to 100%.
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impact of risk factors on the performance of the Nuclear Matrix Protein 22 point of care test for bladder cancer detection
BJUI, 2008Co-Authors: Yair Lotan, Shahrokh F ShariatAbstract:OBJECTIVES To evaluate the impact of different risk factors and symptoms on the performance characteristics of the Nuclear Matrix Protein 22 (NMP22) BladderChek® test (Matritech Inc., Newton, MA, USA) for the detection of bladder cancer, as this is a point-of-care assay that measures NMPs in voided urine. PATIENTS AND METHODS In all, 23 academic, private practice and veterans’ facilities in 10 states prospectively enrolled consecutive patients from September 2001 to May 2002. Participants included 1328 patients at elevated risk of bladder cancer from factors such as a history of smoking or symptoms including haematuria and/or dysuria. Patients at risk of malignancy of the urinary tract provided a voided urine sample for analysis of NMP22 Protein and cytology before cystoscopy. RESULTS Of the 1328 patients: no urinary disease, benign disease and malignancy were found in 545 (41%), 704 (53%) and 79 (6%) patients, respectively. Overall, the positive predictive value (PPV) for detection of bladder cancer was 20.3% (45/222) and negative predictive value (NPV) was 96.9% (1072/1106). The PPV was higher in men (24.0%) than women (13.2%). In men, the PPV increased with smoking (35.4%), gross haematuria (51.2%) and a combination of both factors (70.6%). The impact on the PPV of smoking (9.7%) and gross haematuria (28.6%) was less dramatic in women. The PPV increased from 16.8% in patients aged 65 years. The NPV remained almost always >95% except in men with gross haematuria where it decreased to 77% in smokers and 94% in non-smokers. CONCLUSION The PPV of the BladderChek test improves in patients at higher risk of bladder cancer reaching 77% in men presenting with gross haematuria who are aged >65 years and smoke. The NPV is highest in women aged <65 years, up to 100%.
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variability in the performance of Nuclear Matrix Protein 22 for the detection of bladder cancer
The Journal of Urology, 2006Co-Authors: Shahrokh F Shariat, Yair Lotan, Craig D Zippe, Gerson Ludecke, Hans Boman, Marta Sanchezcarbayo, Martin G Friedrich, Michael Marberger, Ihor S Sawczuk, Roberto CasellaAbstract:Purpose: We assessed variability in the diagnostic performance of NMP22® for detecting recurrence and progression in patients with Ta, T1, and/or CIS transitional cell carcinoma of the bladder in a large international cohort.Materials and Methods: NMP22® voided urine levels were measured in 2,871 patients who underwent office cystoscopy for monitoring previous stage Ta, T1 and/or CIS transitional cell carcinoma at 12 participating institutions.Results: Patient characteristics varied considerably among institutions. Overall 1,045 patients (36.4%) had recurrent transitional cell carcinoma (range across institutions 13.6% to 54.3%). Median NMP22® was 5.5 U/ml (range across institutions 2.5 to 18.8). Of the patients 33.5% had grade III tumors (range across institutions 20.6% to 54.0%) and 22.4% had muscle invasive tumors (range across institutions 3.2% to 38.2%). Area under the ROC curve for bladder TCC detection was 0.735 (95% CI 0.715 to 0.755, range across institutions 0.676 to 0.889). The manufacturer rec...
Roberto Casella - One of the best experts on this subject based on the ideXlab platform.
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urinary cytology and Nuclear Matrix Protein 22 in the detection of bladder cancer recurrence other than transitional cell carcinoma
BJUI, 2008Co-Authors: Georg C Hutterer, Pierre I Karakiewicz, Roberto Casella, Craig D Zippe, Gerson Ludecke, Hans Boman, Marta Sanchezcarbayo, Christine Mian, Martin G Friedrich, Sanaa EissaAbstract:OBJECTIVE: To assess the value of Nuclear Matrix Protein-22 (NMP22), compared with urinary cytology, in predicting the recurrence of bladder cancer that is not transitional cell carcinoma (non-TCC). PATIENTS AND METHODS: We tested the sensitivity, specificity and the predictive accuracy of NMP22 in the context of non-TCC bladder cancer recurrence, and compared it to the performance of urinary cytology. The study group comprised 2687 patients with history of non-muscle-invasive bladder cancer from 10 centres across four continents. RESULTS: The mean patient age was 64.8 years and 75.4% were men; of all patients, 513 (19.1%) had positive urinary cytology, 906 (33.7%) had a positive NMP22 test (
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variability in the performance of Nuclear Matrix Protein 22 for the detection of bladder cancer
The Journal of Urology, 2006Co-Authors: Shahrokh F Shariat, Yair Lotan, Craig D Zippe, Gerson Ludecke, Hans Boman, Marta Sanchezcarbayo, Martin G Friedrich, Michael Marberger, Ihor S Sawczuk, Roberto CasellaAbstract:Purpose: We assessed variability in the diagnostic performance of NMP22® for detecting recurrence and progression in patients with Ta, T1, and/or CIS transitional cell carcinoma of the bladder in a large international cohort.Materials and Methods: NMP22® voided urine levels were measured in 2,871 patients who underwent office cystoscopy for monitoring previous stage Ta, T1 and/or CIS transitional cell carcinoma at 12 participating institutions.Results: Patient characteristics varied considerably among institutions. Overall 1,045 patients (36.4%) had recurrent transitional cell carcinoma (range across institutions 13.6% to 54.3%). Median NMP22® was 5.5 U/ml (range across institutions 2.5 to 18.8). Of the patients 33.5% had grade III tumors (range across institutions 20.6% to 54.0%) and 22.4% had muscle invasive tumors (range across institutions 3.2% to 38.2%). Area under the ROC curve for bladder TCC detection was 0.735 (95% CI 0.715 to 0.755, range across institutions 0.676 to 0.889). The manufacturer rec...
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nomograms including Nuclear Matrix Protein 22 for prediction of disease recurrence and progression in patients with ta t1 or cis transitional cell carcinoma of the bladder
The Journal of Urology, 2005Co-Authors: Shahrokh F Shariat, Roberto Casella, Sanaa Eissa, Craig D Zippe, Gerson Ludecke, Hans Boman, Marta Sanchezcarbayo, Christine Mian, Martin G Friedrich, Hideyuki AkazaAbstract:ABSTRACT Purpose: We developed and validated nomograms that accurately predict disease recurrence and progression in patients with Ta, T1, or CIS transitional cell carcinoma (TCC) of the bladder using a large international cohort. Methods: Univariate and multivariate logistic regression models targeted histologically confirmed disease recurrence, and focused on 2,542 patients with bladder TCC from 10 participating centers. Variables consisted of pre-cystoscopy voided urine Nuclear Matrix Protein 22 (NMP22) assay, urine cytology, age and gender. Resulting nomograms were internally validated with bootstrapping. Nomogram performance was explored graphically with Loess smoothing plots. Results: Overall 957 patients had recurrent TCC. Tumor grade and stage was available for 898 patients, including 24% grade I, 43% grade II, and 33% grade III; 45% stage Ta, 32% T1 and/or CIS, and 23% T2 or greater. Bootstrap corrected predictive accuracy for any TCC recurrence was 0.842; grade III Ta/T1 or CIS was 0.869; and T2 or higher stage TCC of any grade was 0.858. Virtually perfect performance characteristics were observed for the nomograms predicting any TCC recurrence or grade III Ta/T1 or CIS. The nomogram predicting T2 or higher stage TCC overestimated the observed probability for predicted values greater than 45%. Conclusions: We developed and internally validated nomograms that incorporate urinary NMP22, cytology, age and gender to predict with high accuracy the probability of disease recurrence and progression in patients with Ta, T1, and/or CIS bladder TCC. These nomograms could provide a means for individualizing followup in patients with Ta, T1, CIS bladder TCC.
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risk stratification for bladder tumor recurrence stage and grade by urinary Nuclear Matrix Protein 22 and cytology
European Urology, 2004Co-Authors: Shahrokh F Shariat, Roberto Casella, Thomas C Gasser, Tullio Sulser, Frank H Wians, Raheela Ashfaq, Jody Balko, Arthur I SagalowskyAbstract:Abstract Purpose: To test the hypothesis that voided urinary levels of Nuclear Matrix Protein 22 (NMP22) would add to the predictive ability of urine cytology in the diagnosis, staging and grading of bladder transitional cell carcinoma (TCC), and to evaluate the diagnostic performance of different NMP22 cut-points. Materials: NMP22 level and barbotage cytology were evaluated in voided urine specimens collected before cystoscopy from 302 subjects with a history of TCC, 32 subjects with benign urologic pathologies, and 10 healthy volunteers. Results: 180 patients (52%) had bladder TCC. Higher levels of NMP22 and positive cytology were independently associated with an increased risk of TCC, invasive stage, and high grade. The NMP22 value with equal sensitivity and specificity for prediction of bladder cancer was 6.5U/ml; for prediction of grade 3 TCC it was 13.5U/ml; and for prediction of invasive tumor stage it was 17.4U/ml. The NMP22 cut-point of 6.5U/ml outperformed the 10U/ml cut-point in all pathologic stages and grades. The diagnostic sensitivity of the cytology and NMP22 combined was superior across all pathologic stages and grades to that of either marker alone. NMP22 and cytology stratified patients into groups with significantly different risk for TCC presence, invasive stage, and high grade. Conclusions: 6.5U/ml is a robust NMP22 cut-point for bladder cancer surveillance. The diagnostic sensitivities of the combined NMP22 and cytology for TCC presence, stage, and grade were significantly higher than those of single marker alone. The combination of urine cytology and NMP22 could be used to tailor the frequency of cystoscopic follow up.
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the addition of urinary urokinase type plasminogen activator to urinary Nuclear Matrix Protein 22 and cytology improves the detection of bladder cancer
The Journal of Urology, 2003Co-Authors: Shahrokh F Shariat, Roberto Casella, Thomas C Gasser, Mara A Monoski, Tullio Sulser, Seth P LernerAbstract:ABSTRACTPurpose: We have previously reported that urinary urokinase-type plasminogen activator (uPA) and its receptor (uPAR) are elevated in patients with bladder cancer. In the current study we tested the hypothesis that urinary uPA and uPAR would add to the predictive ability of urinary Nuclear Matrix Protein 22 (NMP22) and cytology for the diagnosis of bladder cancer.Materials and Methods: Urinary uPA, uPAR and NMP22 were measured in voided specimens obtained before cystoscopy in 229 consecutive subjects at risk for transitional cell carcinoma (TCC), of whom 122 (53%) were found to have bladder TCC. Bladder washout samples for cytology were also collected in 191 subjects. Associations with TCC were tested by logistic regression. Nonparametric ROC curves were generated and AUCs were compared.Results: Urinary uPA, uPAR and NMP22 were higher in patients with TCC than in controls (p <0.001, 0.016 and <0.001, respectively), while uPA (test for trend p = 0.018) was associated with the risk of TCC after adjus...
Qin Wei - One of the best experts on this subject based on the ideXlab platform.
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sensitive electrochemical immunosensor for detection of Nuclear Matrix Protein 22 based on nh2 sapo 34 supported pd co nanoparticles
Scientific Reports, 2016Co-Authors: Yaoguang Wang, Yong Zhang, Tao Yan, Qin WeiAbstract:A novel sandwich-type electrochemical immunosensor using the new amino group functionalized silicoaluminophosphates molecular sieves (NH2-SAPO-34) supported Pd/Co nanoparticles (NH2-SAPO-34-Pd/Co NPs) as labels for the detection of bladder cancer biomarker Nuclear Matrix Protein-22 (NMP-22) was developed in this work. The reduced graphene oxide-NH (rGO-NH) with good conductivity and large surface area was used to immobilize primary antibody (Ab1). Due to the excellent catalytic activity toward hydrogen peroxide, NH2-SAPO-34-Pd/Co NPs were used as labels and immobilized secondary antibody (Ab2) through adsorption capacity of Pd/Co NPs to Protein. The immunosensor displayed a wide linear range (0.001–20 ng/mL) and low detection limit (0.33 pg/mL). Good reproducibility and stability have showed satisfying results in the analysis of clinical urine samples. This novel and ultrasensitive immunosensor may have the potential application in the detection of different tumor markers.
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gold nanoparticles enhanced electrochemiluminescence of graphite like carbon nitride for the detection of Nuclear Matrix Protein 22
Sensors and Actuators B-chemical, 2014Co-Authors: Tongqian Han, Wei Cao, Qin WeiAbstract:Abstract A novel label-free electrochemiluminescence (ECL) immunosensor was developed for the detection of Nuclear Matrix Protein 22 (NMP 22) based on the graphite-like carbon nitride (g-C3N4) and gold nanoparticles (AuNPs). Graphite-like carbon nitride was firstly combined with AuNPs, which promoted the electrons transfer and enhanced the ECL intensity of g-C3N4 largely. Then, anti-NMP 22 was immobilized onto the electrode through Au–NH2 without the usage of crosslinking agents. The connection between AuNPs and antibody influenced the conductive performance of AuNPs, leading to some decrease of ECL intensity. Then, bovine serum albumin (BSA) solution was dropped on the surface to block nonspecific binding sites. Finally, after the specific immunoreaction between NMP 22 and anti-NMP 22, the ECL intensity decreased further, because the Protein on the electrode hindered the diffusion of luminescent reagents and electrons toward the electrode surface. Therefore, a linear relation between ECL intensity and the logarithm of NMP 22 concentration was obtained from 50.0 pg mL−1 to 2.0 ng/mL (R2 = 0.988) with the detection limit of 10.0 pg mL−1. The proposed ECL immunosensor provides a rapid, simple, and sensitive immunoassay strategy for Protein detection, which might be promising for clinical application.
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a label free electrochemical immunosensor based on au pd ag yolk bimetallic shell nanoparticles and amination graphene for detection of Nuclear Matrix Protein 22
Sensors and Actuators B-chemical, 2014Co-Authors: Yulan Wang, Wei Cao, Qin WeiAbstract:Abstract A sensitive label-free electrochemical immunosensor was developed based on Au@Pd/Ag yolk-bimetallic shell nanoparticles (Au@Pd/Ag NPs) and amination graphene (NH2-GS) for the detection of Nuclear Matrix Protein 22 (NMP22). Au@Pd/Ag NPs were loaded on the surface of NH2-GS (NH2-GS/Au@Pd/Ag) by covalent binding between noble metal NPs and amino groups. A glassy carbon electrode (GCE, 4 mm in diameter) was used as the working electrode for the assembly of NPs and NH2-GS. Due to the large specific surface area and excellent electrical conductivity of NH2-GS/Au@Pd/Ag, the electroactivity of K3[Fe(CN)6] as electron transfer mediator was promoted and the amount of NMP22 antibody immobilized was enhanced significantly. Under optimum conditions, the immunosensor exhibited high selectivity and a wide linear range from 0.01 to 18 ng/mL with a detection limit of 3.3 pg/mL. For real sample analysis, the recovery of standard addition was 97.4–99.3% and the relative standard deviation (RSD) was 4.83–7.21%, which indicated that the proposed method exhibited great potential as a reliable technique for the determination of NMP22 for clinical research and diagnostic applications.
Stephen P Goff - One of the best experts on this subject based on the ideXlab platform.
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Nuclear Matrix Protein matrin 3 is a regulator of zap mediated retroviral restriction
Retrovirology, 2015Co-Authors: Angela Erazo, Stephen P GoffAbstract:Matrin 3 is a Nuclear Matrix Protein involved in multiple Nuclear processes. In HIV-1 infection, Matrin 3 serves as a Rev cofactor important for the cytoplasmic accumulation of HIV-1 transcripts. ZAP is a potent host restriction factor of multiple viruses including retroviruses HIV-1 and MoMuLV. In this study we sought to further characterize Matrin 3 functions in the regulation of HIV gene expression. Here we describe a function for Matrin 3 as a negative regulator of the ZAP-mediated restriction of retroviruses. Mass spectrometry analysis of Matrin 3-associated Proteins uncovered interactions with Proteins of the ZAP degradation complex, DDX17 and EXOSC3. Coimmunoprecipitation studies confirmed Matrin 3 associations with DDX17, EXOSC3 and ZAP, in a largely RNA-dependent manner, indicating that RNA is mediating the Matrin 3 interactions with these components of the ZAP degradation complex. Silencing Matrin 3 expression caused a remarkably enhanced ZAP-driven inhibition of HIV-1 and MoMuLV luciferase reporter viruses. This effect was shared with additional Nuclear Matrix Proteins. ZAP targets multiply-spliced HIV-1 transcripts, but in the context of Matrin 3 suppression, this ZAP restriction was broadened to unspliced and multiply-spliced RNAs. Here we reveal an unprecedented role for a Nuclear Matrix Protein, Matrin 3, in the regulation of ZAP’s antiretroviral activity. Suppressing Matrin 3 powers a heightened and broader ZAP restriction of HIV-1 gene expression. This study suggests that this ZAP regulatory mechanism is shared with additional Nuclear Matrix Proteins.
