Urho formation of the Mabei slope zone in Junggar basin had a stable structure and developed fan ... more Urho formation of the Mabei slope zone in Junggar basin had a stable structure and developed fan delta sedimentary system during the deposition process. Distributary channel and fan River constitute the sedimentary framework Urho formation. The research on diagenesis of Urho reservoir based on a large number of thin section and core analysis is conducted by different means of experiments such as scanning electron microscopy, X-ray diffractions and cathode luminescence, etc. The results show that the influence of diagenesis on the reservoir is duality. The effect of compaction (pressure solution) in the study layer is mainly controlled by the difference of vertical burial. The calcite cementation is mainly formed at the early stage of burial. Since the late-early diagenetic stage to the intermediate diagenetic stage, large scale of acidic dissolution and alteration occurred in Urho formation. Acid dissolution mainly exists in widely developed volcanic rocks and zeolite cements, which...
With the help of core and logging data, laboratory test data, by using classified evaluation of g... more With the help of core and logging data, laboratory test data, by using classified evaluation of glutenite based on classification of sedimentation, subdividing the glutenite facies into tractive-current glutenite and gravity- current glutenite. The tractive-current glutenite is mainly a set of fan channel deposits and underwater distributary channel deposits. Gravity-current glutenite is a set of debris flow deposition and debris flow deposition. Predictive results are found to be high accuracy by first time using glutenite logging identification technology, pseudo-sonic wave impedance technique. The result of research shows that the difference of sedimentation and conglomerate is the main factor controlling the reservoir physical properties and productivity in Mabei area. The physical and oily and oil test production are mainly controlled by the sedimentary genesis of the glutenite reservoir and less by diagenesis modification. The results show that the development of tractive-curr...
The present study aimed to investigate the role and underlying mechanisms of long non-coding RNA ... more The present study aimed to investigate the role and underlying mechanisms of long non-coding RNA (lncRNA) muscleblind-like 1 antisense RNA 1 (MBNL1-AS1) in the progression of Prostate cancer (PCa). MBNL1-AS1 and microRNA (miR)-181a-5p expression in PCa tissues and several human PCa cell lines were analyzed, respectively, using StarBasev3.0 project and RT-qPCR assay. After MBNL1-AS1 overexpression, cell proliferation, invasion and migration were, respectively, evaluated using CCK-8, colony formation, transwell and wound healing assays. Dual luciferase assay were used for analysis of the interactions among MBNL1-AS1, miR-181a-5p, and phosphatase and tensin homolog (PTEN). Subsequently, the expression of PTEN and proteins in PI3K/AKT/mTOR signaling was examined using western blot analysis after transfection with miR-181a-5p mimic. The rescue assays were performed to investigate the effects of MBNL1-AS1 and miR-181a-5p on the functions of PCa cells and the expression of PTEN/PI3K/AKT/mTOR signaling by co-transfection with MBNL1-AS1 plasmid and miR-181a-5p mimic. Results indicated that MBNL1-AS1 was conspicuously downregulated while miR-181a-5p upregulating in PCa tissues and cell lines. MBNL1-AS1 overexpression decreased the abilities of cell proliferation, invasion, and migration. Further study revealed that MBNL1-AS1 acted as a sponge for miR-181a-5p and positively regulated PTEN by a sponge effect. Additionally, rescue assays proved that the effect of MBNL1-AS1-upregulation on the proliferation, invasion, and migration of PCa cells was dependent on miR-181a-5p. Furthermore, miR-181a-5p overexpression counteracted the expression of PTEN and proteins in PI3K/AKT/mTOR signaling exerted by MBNL1-AS1-upregulation in PCa cells. This study suggests that MBNL1-AS1 inhibits the progression of PCa via sponging miR-181a-5p and regulating PTEN/PI3K/AKT/mTOR pathway.
2009 3rd International Conference on Bioinformatics and Biomedical Engineering, 2009
Nylon membranes are first hydrolyed with dilute HCl and then treated with chitosan before being u... more Nylon membranes are first hydrolyed with dilute HCl and then treated with chitosan before being used as the affinity carrier. Papain as a ligand has been immobilized on the activated membranes with glutaraldehyde as a crosslinking agent. The factors involving with the activity of immobilized papain, such as concentration of glutaraldehyde, pH, temperature, reaction time, and the amount of added papain have been studied. The results show that the optimum conditions for the preparation of immobilized papain nylon affinity membranes are as follows: pH 9.0, 0.5% glutaraldehyde solution, the concentration of added papain is 10 mg/ml, the reaction time is 6 h at 45degC. The nylon membrane prepared is then used to purify cystatin from potato juice, and shows that they are high efficiency affinity membranes base for cystatin separation.
Abstract : The goal of this research was to demonstrate the feasibility of using flow cytometry t... more Abstract : The goal of this research was to demonstrate the feasibility of using flow cytometry to detect prostate cancer cells in seminal fluid as an important step toward developing a new test for the non-invasive diagnosis of prostate cancer. Results obtained during the first 12 months of the project were promising. Tragically, the Principal Investigator, Dr. Gerald P. Murphy, died in the l6th month of the project period. After Dr. Murphy's death, a reassessment of the technique used to prepare seminal fluid samples for flow cytometry showed that these methods were not adequate for the repeatable detection of prostate cancer cells. As a result, a revised Statement of Work was approved which focused on developing a more robust method of epithelial cell detection in seminal fluid that would overcome sperm interference. Magnetic bead separation improved the recovery of prostate cancer cells from semen, but failed to consistently produce sufficient yield of epithelial cells from patient samples. Fixation of seminal fluid samples, a prerequisite for the practical application of this assay, significantly reduced prostate cell recovery using magnetic bead separation. In summary, our work failed to define conditions necessary to establish seminal fluid analysis using flow cytometry as a reliable assay for the detection of prostate cancer.
Journal of Shanghai Jiaotong University (Science), 2014
With more successful applications of advanced medical imaging technologies in clinical diagnosis,... more With more successful applications of advanced medical imaging technologies in clinical diagnosis, various analytic discriminant approaches, by seeking the imaging based characteristics of a given disease to achieve automatic diagnosis, gain greater attention in the medical community. However the existing computer-aided discriminant procedures for Alzheimer's disease (AD) are yet to be improved for better identifying patients with mild cognitive impairment (MCI) from those with AD and those who are cognitively normal. In this work we present a computer assisted diagnosis approach by first statistically extracting characteristics from whole brain 2-deoxy-2-(18 F)fluoro-D-glucose positron emission tomography (18 F-FDG PET) images, and then using support vector machines for classification. Evaluations of the proposed procedure with patient data exhibit satisfactory accuracies in distinguishing AD from its early stage MCI, and normal controls.
Serum prostate-specific antigen is credited with dramatic advances in the early detection, screen... more Serum prostate-specific antigen is credited with dramatic advances in the early detection, screening, and management of men with prostatic carcinoma. There has been more than a twofold increase in the number of men diagnosed during the last decade, and prostate cancer has emerged as the most common non-skin cancer and the second leading cause of cancer death in men. This report summarizes the history and current status of prostate-specific antigen and other serum markers, incorporating consensus opinions from the Second International Consultation on Prostate Cancer held in Paris in June 1999.
The human fibroblast activation protein (FAP), defined by monoclonal antibody F19, is expressed i... more The human fibroblast activation protein (FAP), defined by monoclonal antibody F19, is expressed in vivo in reactive stromal fibroblasts of epithelial cancers, subsets of bone and soft tissue sarcomas, and granulation tissue of healing wounds. FAP is generally absent from the stroma of benign epithelial tumors and normal adult tissues. In vitro FAP induction is observed in proliferating cultured fibroblasts and in melanocytes grown with fibroblast growth factor and phorbol ester. In the present study, we show that fibroblast and melanocyte FAP is a cell surface protein comprising noncovalently linked M(r) 95,000 (p95) and M(r) 105,000 (p105) subunits. In contrast, cultured sarcoma and melanoma cell lines express only p95 or are FAP negative. Immunoblot experiments show that p95, but not p105, carries the epitope defined by monoclonal antibody F19. Furthermore, peptide maps of purified p95 and p105 differ, suggesting that they may be distinct gene products. Loss of FAP or a change fro...
We examined expression of prostate-specific membrane antigen (PSM) mRNA in normal prostate using ... more We examined expression of prostate-specific membrane antigen (PSM) mRNA in normal prostate using reverse transcription-PCR and sequencing. An alternatively spliced variant, PSM', along with the previously described PSM form, was found in normal prostate. PSM' cDNA is shorter (2387 nucleotides) than PSM (2653 nucleotides). The cDNAs are identical except for a 266-nucleotide region near the 5' end of PSM cDNA (nucleotide 114-380) that is absent from PSM'. This deleted region includes the translation initiation codon and codons for the putative transmembrane domain of PSM. Thus, PSM' RNA codes for a protein that has no apparent signal sequence. We verified the existence of spliced mRNA variants in human primary tissue specimens by RNase protection assay. In LNCaP human prostatic cancer cells and in primary prostate tumors, PSM is the dominant form. In contrast, normal human prostate expressed more PSM' than PSM. Benign prostatic hypertrophy samples showed about ...
A highly sensitive nested reverse transcriptase-PCR assay, with primers derived from the prostate... more A highly sensitive nested reverse transcriptase-PCR assay, with primers derived from the prostate-specific antigen (PSA) and prostate-specific membrane antigen (PSM) cDNA sequences, has been used to detect occult hematogenous micrometastatic prostate cells. In 77 patients with prostate cancer, PSM and PSA primers detected circulating prostate cells in 48 (62.3%) and 7 (9.1%) patients, respectively. In treated stage D disease patients, PSM primers detected cells in 16 of 24 patients (66.7%), while PSA primers detected cells in 6 of 24 (25%). In post-radical prostectomy patients with negative serum PSA values, PSM primers detected metastases in 21 of 31 patients (67.7%), whereas PSA primers detected cells in only 1 of 33 (3.0%), indicating that micrometastatic spread may be a relatively early event in prostate cancer. The analysis of 40 individuals without known prostate cancer provides evidence that this assay is highly specific and suggests that PSM expression may predict the develo...
The HMI1 gene encodes a DNA helicase that localizes to the mitochondria and is required for maint... more The HMI1 gene encodes a DNA helicase that localizes to the mitochondria and is required for maintenance of the mitochondrial DNA (mtDNA) genome of Saccharomyces cerevisiae. Identified based on its homology with E. coli uvrD, the HMI1 gene product, Hmi1p, has been presumed to be involved in the replication of the 80 kb linear S. cerevisiae mtDNA genome. Here we report the purification of Hmi1p to apparent homogeneity and provide a characterization of the helicase reaction and the ATPase reaction with regard to NTP preference, divalent cation preference and the stimulatory effects of different nucleic acids on Hmi1p-catalysed ATPase activity. Genetic complementation assays indicate that mitochondrial localization of Hmi1p is essential for its role in mtDNA metabolism. The helicase activity, however, is not essential. Point mutants that lack ATPase/helicase activity partially complement a strain lacking Hmi1p. We suggest several possible roles for Hmi1p in mtDNA metabolism.
Objective. An ongoing study at the Memorial Sloan-Kettering Cancer Center assessed the effectiven... more Objective. An ongoing study at the Memorial Sloan-Kettering Cancer Center assessed the effectiveness of androgen deprivation therapy (ADT) prior to surgical removal of the prostate. In this report, we evaluate the effectiveness of ADT on systemic disease by monitoring the presence or absence of circulating prostatic epithelial cells using a reverse transcription polymerase chain reaction (RT-PCR) assay for prostatic-specific membrane antigen (PSM). Methods. PSM RT-PCR was performed on a total of 38 prostate cancer patients. There were 12 pT2 patients in the ADT group and 10 patients in the control pT2 group and 5 pT3 patients in the ADT group and 1 1 pT3 patients in the control group. Results. For pT2 patients, 2 of the 12 patients (17%) were positive for circulating prostatic cells during androgen deprivation therapy but before radical retroprostatectomy (RRP). Within a 6-month period after RRP, 3 of 12 patients (25%) were positive. For the period between the 7th and 12th month after RRP, 6 of 12 patients (50%) were positive. For the period 12-36 months after RRP, 2 of the 12 patients (17%) remained positive for circulating prostatic cells. In contrast, the pT2 control group had higher positive rates in comparable periods: 4 of 10 patients (40%) were positive prior to surgery; 6 of 10 patients (60%) were positive during the 6 months following surgery. For the period between the 7th and 12th month following surgery, 4 of 7 patients (57%) were positive for PSM. Finally, 3 of 6 patients (50%) were positive for the period longer than 12 months. Regarding patients who have extraprostatic disease (stage pT3), the ADT group had a lower rate of circulating PSM positive cells. Before RRP and during androgen deprivation therapy, 1 out of 5 patients (20%) in the ADT group were positive as compared to 4 out of 1 1 patients for the control group. Within a 6-month period after RRP, the ADT group had 4 out of 9 (44%) patients positive for PSM as compared to 9 of 1 1 (82%) for the control group. For the period between the 7th and 12th months postsurgery, 1 of 5 patients (20%) of the ADT group were positive as compared to 4 of 7 (57%) of the control patients. Conclusions. These results indicate that patients with pT2 and pT3 lesions who receive neoadjuvant ADT are less likely to have circulating tumor cells detected compared to a control group both prior to and after surgery. In addition, irrespective of ADT or control group, there were increases in the detection of circulating tumor cells in the period after RRP, and this rise gradually decreased, suggesting that surgical manipulation may cause hematogenous dissemination of tumor cells and that ADT reduces such dissemination of tumor cells. Overall, these results indicate that the use of neoadjuvant ADT decreases the number of circulating prostatic cells. These data represent the initial results of an ongoing study. As additional patients are added to the studies, attempts to correlate PSM positivity and serum PSAvalues postoperatively, recurrence, and margin positivity will be made.
This investigation attempts to study and optimize the affinity partitioning conditions of papain ... more This investigation attempts to study and optimize the affinity partitioning conditions of papain in an aqueous two-phase system (ATPS). Reactive Red 120 was added to the ATPS as a free affinity dye ligand and five factors (pH, PEG concentration, ammonium sulfate, sodium chloride and dye) that influence the papain partitioning were analyzed using response surface methodology. The optimum conditions were determined as PEG 0.102 g/ml, ammonium sulfate 0.17 g/ml, sodium chloride 25 mg/ml, Reactive Red 120 4.5 g/ml and pH 8.0. It was observed that sodium chloride increased papain partitioning and Reactive Red 120 had little effect on partitioning. The optimal condition gave a K U value for papain partitioning of 1.92 with a yield of 50.6%.
Urho formation of the Mabei slope zone in Junggar basin had a stable structure and developed fan ... more Urho formation of the Mabei slope zone in Junggar basin had a stable structure and developed fan delta sedimentary system during the deposition process. Distributary channel and fan River constitute the sedimentary framework Urho formation. The research on diagenesis of Urho reservoir based on a large number of thin section and core analysis is conducted by different means of experiments such as scanning electron microscopy, X-ray diffractions and cathode luminescence, etc. The results show that the influence of diagenesis on the reservoir is duality. The effect of compaction (pressure solution) in the study layer is mainly controlled by the difference of vertical burial. The calcite cementation is mainly formed at the early stage of burial. Since the late-early diagenetic stage to the intermediate diagenetic stage, large scale of acidic dissolution and alteration occurred in Urho formation. Acid dissolution mainly exists in widely developed volcanic rocks and zeolite cements, which...
With the help of core and logging data, laboratory test data, by using classified evaluation of g... more With the help of core and logging data, laboratory test data, by using classified evaluation of glutenite based on classification of sedimentation, subdividing the glutenite facies into tractive-current glutenite and gravity- current glutenite. The tractive-current glutenite is mainly a set of fan channel deposits and underwater distributary channel deposits. Gravity-current glutenite is a set of debris flow deposition and debris flow deposition. Predictive results are found to be high accuracy by first time using glutenite logging identification technology, pseudo-sonic wave impedance technique. The result of research shows that the difference of sedimentation and conglomerate is the main factor controlling the reservoir physical properties and productivity in Mabei area. The physical and oily and oil test production are mainly controlled by the sedimentary genesis of the glutenite reservoir and less by diagenesis modification. The results show that the development of tractive-curr...
The present study aimed to investigate the role and underlying mechanisms of long non-coding RNA ... more The present study aimed to investigate the role and underlying mechanisms of long non-coding RNA (lncRNA) muscleblind-like 1 antisense RNA 1 (MBNL1-AS1) in the progression of Prostate cancer (PCa). MBNL1-AS1 and microRNA (miR)-181a-5p expression in PCa tissues and several human PCa cell lines were analyzed, respectively, using StarBasev3.0 project and RT-qPCR assay. After MBNL1-AS1 overexpression, cell proliferation, invasion and migration were, respectively, evaluated using CCK-8, colony formation, transwell and wound healing assays. Dual luciferase assay were used for analysis of the interactions among MBNL1-AS1, miR-181a-5p, and phosphatase and tensin homolog (PTEN). Subsequently, the expression of PTEN and proteins in PI3K/AKT/mTOR signaling was examined using western blot analysis after transfection with miR-181a-5p mimic. The rescue assays were performed to investigate the effects of MBNL1-AS1 and miR-181a-5p on the functions of PCa cells and the expression of PTEN/PI3K/AKT/mTOR signaling by co-transfection with MBNL1-AS1 plasmid and miR-181a-5p mimic. Results indicated that MBNL1-AS1 was conspicuously downregulated while miR-181a-5p upregulating in PCa tissues and cell lines. MBNL1-AS1 overexpression decreased the abilities of cell proliferation, invasion, and migration. Further study revealed that MBNL1-AS1 acted as a sponge for miR-181a-5p and positively regulated PTEN by a sponge effect. Additionally, rescue assays proved that the effect of MBNL1-AS1-upregulation on the proliferation, invasion, and migration of PCa cells was dependent on miR-181a-5p. Furthermore, miR-181a-5p overexpression counteracted the expression of PTEN and proteins in PI3K/AKT/mTOR signaling exerted by MBNL1-AS1-upregulation in PCa cells. This study suggests that MBNL1-AS1 inhibits the progression of PCa via sponging miR-181a-5p and regulating PTEN/PI3K/AKT/mTOR pathway.
2009 3rd International Conference on Bioinformatics and Biomedical Engineering, 2009
Nylon membranes are first hydrolyed with dilute HCl and then treated with chitosan before being u... more Nylon membranes are first hydrolyed with dilute HCl and then treated with chitosan before being used as the affinity carrier. Papain as a ligand has been immobilized on the activated membranes with glutaraldehyde as a crosslinking agent. The factors involving with the activity of immobilized papain, such as concentration of glutaraldehyde, pH, temperature, reaction time, and the amount of added papain have been studied. The results show that the optimum conditions for the preparation of immobilized papain nylon affinity membranes are as follows: pH 9.0, 0.5% glutaraldehyde solution, the concentration of added papain is 10 mg/ml, the reaction time is 6 h at 45degC. The nylon membrane prepared is then used to purify cystatin from potato juice, and shows that they are high efficiency affinity membranes base for cystatin separation.
Abstract : The goal of this research was to demonstrate the feasibility of using flow cytometry t... more Abstract : The goal of this research was to demonstrate the feasibility of using flow cytometry to detect prostate cancer cells in seminal fluid as an important step toward developing a new test for the non-invasive diagnosis of prostate cancer. Results obtained during the first 12 months of the project were promising. Tragically, the Principal Investigator, Dr. Gerald P. Murphy, died in the l6th month of the project period. After Dr. Murphy's death, a reassessment of the technique used to prepare seminal fluid samples for flow cytometry showed that these methods were not adequate for the repeatable detection of prostate cancer cells. As a result, a revised Statement of Work was approved which focused on developing a more robust method of epithelial cell detection in seminal fluid that would overcome sperm interference. Magnetic bead separation improved the recovery of prostate cancer cells from semen, but failed to consistently produce sufficient yield of epithelial cells from patient samples. Fixation of seminal fluid samples, a prerequisite for the practical application of this assay, significantly reduced prostate cell recovery using magnetic bead separation. In summary, our work failed to define conditions necessary to establish seminal fluid analysis using flow cytometry as a reliable assay for the detection of prostate cancer.
Journal of Shanghai Jiaotong University (Science), 2014
With more successful applications of advanced medical imaging technologies in clinical diagnosis,... more With more successful applications of advanced medical imaging technologies in clinical diagnosis, various analytic discriminant approaches, by seeking the imaging based characteristics of a given disease to achieve automatic diagnosis, gain greater attention in the medical community. However the existing computer-aided discriminant procedures for Alzheimer's disease (AD) are yet to be improved for better identifying patients with mild cognitive impairment (MCI) from those with AD and those who are cognitively normal. In this work we present a computer assisted diagnosis approach by first statistically extracting characteristics from whole brain 2-deoxy-2-(18 F)fluoro-D-glucose positron emission tomography (18 F-FDG PET) images, and then using support vector machines for classification. Evaluations of the proposed procedure with patient data exhibit satisfactory accuracies in distinguishing AD from its early stage MCI, and normal controls.
Serum prostate-specific antigen is credited with dramatic advances in the early detection, screen... more Serum prostate-specific antigen is credited with dramatic advances in the early detection, screening, and management of men with prostatic carcinoma. There has been more than a twofold increase in the number of men diagnosed during the last decade, and prostate cancer has emerged as the most common non-skin cancer and the second leading cause of cancer death in men. This report summarizes the history and current status of prostate-specific antigen and other serum markers, incorporating consensus opinions from the Second International Consultation on Prostate Cancer held in Paris in June 1999.
The human fibroblast activation protein (FAP), defined by monoclonal antibody F19, is expressed i... more The human fibroblast activation protein (FAP), defined by monoclonal antibody F19, is expressed in vivo in reactive stromal fibroblasts of epithelial cancers, subsets of bone and soft tissue sarcomas, and granulation tissue of healing wounds. FAP is generally absent from the stroma of benign epithelial tumors and normal adult tissues. In vitro FAP induction is observed in proliferating cultured fibroblasts and in melanocytes grown with fibroblast growth factor and phorbol ester. In the present study, we show that fibroblast and melanocyte FAP is a cell surface protein comprising noncovalently linked M(r) 95,000 (p95) and M(r) 105,000 (p105) subunits. In contrast, cultured sarcoma and melanoma cell lines express only p95 or are FAP negative. Immunoblot experiments show that p95, but not p105, carries the epitope defined by monoclonal antibody F19. Furthermore, peptide maps of purified p95 and p105 differ, suggesting that they may be distinct gene products. Loss of FAP or a change fro...
We examined expression of prostate-specific membrane antigen (PSM) mRNA in normal prostate using ... more We examined expression of prostate-specific membrane antigen (PSM) mRNA in normal prostate using reverse transcription-PCR and sequencing. An alternatively spliced variant, PSM', along with the previously described PSM form, was found in normal prostate. PSM' cDNA is shorter (2387 nucleotides) than PSM (2653 nucleotides). The cDNAs are identical except for a 266-nucleotide region near the 5' end of PSM cDNA (nucleotide 114-380) that is absent from PSM'. This deleted region includes the translation initiation codon and codons for the putative transmembrane domain of PSM. Thus, PSM' RNA codes for a protein that has no apparent signal sequence. We verified the existence of spliced mRNA variants in human primary tissue specimens by RNase protection assay. In LNCaP human prostatic cancer cells and in primary prostate tumors, PSM is the dominant form. In contrast, normal human prostate expressed more PSM' than PSM. Benign prostatic hypertrophy samples showed about ...
A highly sensitive nested reverse transcriptase-PCR assay, with primers derived from the prostate... more A highly sensitive nested reverse transcriptase-PCR assay, with primers derived from the prostate-specific antigen (PSA) and prostate-specific membrane antigen (PSM) cDNA sequences, has been used to detect occult hematogenous micrometastatic prostate cells. In 77 patients with prostate cancer, PSM and PSA primers detected circulating prostate cells in 48 (62.3%) and 7 (9.1%) patients, respectively. In treated stage D disease patients, PSM primers detected cells in 16 of 24 patients (66.7%), while PSA primers detected cells in 6 of 24 (25%). In post-radical prostectomy patients with negative serum PSA values, PSM primers detected metastases in 21 of 31 patients (67.7%), whereas PSA primers detected cells in only 1 of 33 (3.0%), indicating that micrometastatic spread may be a relatively early event in prostate cancer. The analysis of 40 individuals without known prostate cancer provides evidence that this assay is highly specific and suggests that PSM expression may predict the develo...
The HMI1 gene encodes a DNA helicase that localizes to the mitochondria and is required for maint... more The HMI1 gene encodes a DNA helicase that localizes to the mitochondria and is required for maintenance of the mitochondrial DNA (mtDNA) genome of Saccharomyces cerevisiae. Identified based on its homology with E. coli uvrD, the HMI1 gene product, Hmi1p, has been presumed to be involved in the replication of the 80 kb linear S. cerevisiae mtDNA genome. Here we report the purification of Hmi1p to apparent homogeneity and provide a characterization of the helicase reaction and the ATPase reaction with regard to NTP preference, divalent cation preference and the stimulatory effects of different nucleic acids on Hmi1p-catalysed ATPase activity. Genetic complementation assays indicate that mitochondrial localization of Hmi1p is essential for its role in mtDNA metabolism. The helicase activity, however, is not essential. Point mutants that lack ATPase/helicase activity partially complement a strain lacking Hmi1p. We suggest several possible roles for Hmi1p in mtDNA metabolism.
Objective. An ongoing study at the Memorial Sloan-Kettering Cancer Center assessed the effectiven... more Objective. An ongoing study at the Memorial Sloan-Kettering Cancer Center assessed the effectiveness of androgen deprivation therapy (ADT) prior to surgical removal of the prostate. In this report, we evaluate the effectiveness of ADT on systemic disease by monitoring the presence or absence of circulating prostatic epithelial cells using a reverse transcription polymerase chain reaction (RT-PCR) assay for prostatic-specific membrane antigen (PSM). Methods. PSM RT-PCR was performed on a total of 38 prostate cancer patients. There were 12 pT2 patients in the ADT group and 10 patients in the control pT2 group and 5 pT3 patients in the ADT group and 1 1 pT3 patients in the control group. Results. For pT2 patients, 2 of the 12 patients (17%) were positive for circulating prostatic cells during androgen deprivation therapy but before radical retroprostatectomy (RRP). Within a 6-month period after RRP, 3 of 12 patients (25%) were positive. For the period between the 7th and 12th month after RRP, 6 of 12 patients (50%) were positive. For the period 12-36 months after RRP, 2 of the 12 patients (17%) remained positive for circulating prostatic cells. In contrast, the pT2 control group had higher positive rates in comparable periods: 4 of 10 patients (40%) were positive prior to surgery; 6 of 10 patients (60%) were positive during the 6 months following surgery. For the period between the 7th and 12th month following surgery, 4 of 7 patients (57%) were positive for PSM. Finally, 3 of 6 patients (50%) were positive for the period longer than 12 months. Regarding patients who have extraprostatic disease (stage pT3), the ADT group had a lower rate of circulating PSM positive cells. Before RRP and during androgen deprivation therapy, 1 out of 5 patients (20%) in the ADT group were positive as compared to 4 out of 1 1 patients for the control group. Within a 6-month period after RRP, the ADT group had 4 out of 9 (44%) patients positive for PSM as compared to 9 of 1 1 (82%) for the control group. For the period between the 7th and 12th months postsurgery, 1 of 5 patients (20%) of the ADT group were positive as compared to 4 of 7 (57%) of the control patients. Conclusions. These results indicate that patients with pT2 and pT3 lesions who receive neoadjuvant ADT are less likely to have circulating tumor cells detected compared to a control group both prior to and after surgery. In addition, irrespective of ADT or control group, there were increases in the detection of circulating tumor cells in the period after RRP, and this rise gradually decreased, suggesting that surgical manipulation may cause hematogenous dissemination of tumor cells and that ADT reduces such dissemination of tumor cells. Overall, these results indicate that the use of neoadjuvant ADT decreases the number of circulating prostatic cells. These data represent the initial results of an ongoing study. As additional patients are added to the studies, attempts to correlate PSM positivity and serum PSAvalues postoperatively, recurrence, and margin positivity will be made.
This investigation attempts to study and optimize the affinity partitioning conditions of papain ... more This investigation attempts to study and optimize the affinity partitioning conditions of papain in an aqueous two-phase system (ATPS). Reactive Red 120 was added to the ATPS as a free affinity dye ligand and five factors (pH, PEG concentration, ammonium sulfate, sodium chloride and dye) that influence the papain partitioning were analyzed using response surface methodology. The optimum conditions were determined as PEG 0.102 g/ml, ammonium sulfate 0.17 g/ml, sodium chloride 25 mg/ml, Reactive Red 120 4.5 g/ml and pH 8.0. It was observed that sodium chloride increased papain partitioning and Reactive Red 120 had little effect on partitioning. The optimal condition gave a K U value for papain partitioning of 1.92 with a yield of 50.6%.
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