Anthony davies - Academia.edu (original) (raw)

Papers by Anthony davies

European Journal of Cancer Supplements

Background: Tissue microarray analyses of human breast carcinomas have linked decreased annexin I... more Background: Tissue microarray analyses of human breast carcinomas have linked decreased annexin I (ANXA-1) expression with breast cancer progression [1]. ANXA-1, a calcium-and phospholipid-binding protein, has been implicated in A549 lung cancer cell growth and apoptosis [2]. We sought to establish the effect of ANXA-1 protein knockdown on cell cycle regulation in the estrogen receptor (ER)-positive MCF-7 cells and ERnegative MDA-MB-231 breast tumour cell lines. Materials & methods: MCF-7 and MDA-MB-231 cells were transfected with either ANXA-1 targeting stealth RNAi (siRNA) or its scrambled control sequence. After 24 hours, cells were stimuated with 5% FC. Cell lysates were obtained 24 and 48 hours post-transfection for analysis of ANXA-1 protein levels. Enumeration of viable cells was performed after a 48 hour incubation with FCS and cell cycle regulation was determined by flow cytometry of propidium iodide stained cells. Results: At 24 and 48 hours post-transfection, ANXA-1 protein levels were significantly decreased by 28% and 55% respectively in MCF-7 cells. The reduction of ANXA-1 in MDA-MB-231 cells at 48 hours after transfection was 35% (p < 0.05). ANXA-1 knockdown reduced basal and FCS-induced increases in MCF-7 cell number, accompanied, at 48 by significant reduction in G2/M phase cells. Neither basal nor FCS-induced increases in cell number were influenced by ANXA-1 siRNA in the ERnegative MDA-MB-231 cell line. Conclusions: These observations implicate ANXA-1 in the FCS-induced proliferation of MCF-7 cells. 1. Shen, D., et al., Decreased expression of annexin A1 is correlated with breast cancer development and progression as determined by a tissue microarray analysis.

Journal of Thoracic Oncology

Background: Bevacizumab (BEV), a humanized recombinant monoclonal antibody that targets vascular ... more Background: Bevacizumab (BEV), a humanized recombinant monoclonal antibody that targets vascular endothelial growth factor, is widely used in cancer treatment. BEV treatment is generally well-tolerated, however, patients who are treated with BEV have an increased risk of developing systemic adverse events, such as hypertension and proteinuria. We generally use angiotensin Iconverting enzyme inhibitors (ACEIs) and angiotensin II type-1 receptor blockers (ARBs) as a treatment of these adverse events. This group of drugs has been found to reduce proteinuria and prevent the development of glomeruloscrelosis. Additionally, since angiotensin II stimulates neovascularization, and thus act as a growth factor for cancer, previous studies have suggested that ACEIs and ARBs might decrease tumor growth and tumor-associated angiogenesis and inhibit metastasis. In this study, we aimed to investigate the correlation between proteinuria/hypertension and clinical outcome, and between ACEIs/ARBs use and clinical outcome in lung cancer patients treated with BEV-based chemotherapy. Methods: We retrospectively reviewed medical charts of patients at Kobe Red Cross Hospital and Kurashiki Central Hospital between November 2009 and February 2014. All patients were treated with BEV in combination with standard chemotherapy, such as pemetrexed plus platinum doublet, as first or second-line treatment. Cox regression analysis was performed to identify factors associated with antitumor response, overall survival (OS), and progression free survival (PFS). Results: Among 122 patients, almost 30 patients were excluded because of diabetes mellitus and use of .BEV in late lines. Ninety-nine patients were included in the analysis set. Median treated BEV cycles were 7 cycles. Median OS and median PFS were 13.0 months and 6.2 months, respectively. During the study, proteinuria was found in 56 patients (57%). Hypertension was found in 81 patients (82%). Thirty-nine patients (39%) were treated with ACEIs/ ARBs, 26 patients with other drugs. Univariate analysis showed that younger age, BEV cycles, and ACEIs/ARBs use were significantly associated with longer PFS, not OS. Younger age and BEV cycles were associated with PFS (P ¼ .030 and < 0.001, respectively) on multivariate analysis, however, proteinuria, hypertension, and ACEIs/ARBs use were not. There was however a trend for the correlation between ACEIs/ ARBs use and anti-tumor response (p ¼ 0.082). Conclusion: Our results suggest that BEV-related proteinuria and hypertension were not prognostic markers for lung cancer patients treated with BEV-based chemotherapy. Further accumulation of patients is needed to assess the potential effect of ACEIs/ARBs on anti-tumor effect and PFS.

Journal of Thoracic Oncology

Background: Stathmin 1 is a cytosolic phosphoprotein that plays a crucial role in the control of ... more Background: Stathmin 1 is a cytosolic phosphoprotein that plays a crucial role in the control of cellular division and proliferation by regulating microtubule dynamics. In addition, Stathmin1 is associated with tumor growth and progression. Our study aimed to determine differences in overall (OS) and disease free survival (DFS) in patients with non-small lung cancer (NSCLC) stratified by STMN1 tumor expression.

Nature Reviews Drug Discovery, 2016

The common and persistent failures to translate promising preclinical drug candidates into clinic... more The common and persistent failures to translate promising preclinical drug candidates into clinical success highlight the limited effectiveness of disease models currently used in drug discovery. An apparent reluctance to explore and adopt alternative cell-and tissuebased model systems, coupled with a detachment from clinical practice during assay validation, contributes to ineffective translational research. To help address these issues and stimulate debate, here we propose a set of principles to facilitate the definition and development of disease-relevant assays, and we discuss new opportunities for exploiting the latest advances in cell-based assay technologies in drug discovery, including induced pluripotent stem cells, 3D co-culture and organ-on-a-chip systems, complemented by advances in single-cell imaging and gene editing technologies. Funding to support precompetitive, multidisciplinary collaboration to develop novel preclinical models and cell-based screening technologies could have a key role in improving their clinical relevance, and ultimately increase clinical success rates [Au: edits to shorten OK?].

Oncotarget, 2014

Head and neck cancers are the seventh most common cancer globally. Approximately 50% of the patie... more Head and neck cancers are the seventh most common cancer globally. Approximately 50% of the patients die within 5 years. Current diagnostic methods include clinical assessment, imaging and tissue biopsy [1]. When metastases are clinically evident, treatment is palliative in nature only. There are currently no methods to predict which patients with a higher disease burden will develop metastases. The ability to do so would potentially lend itself to treatment escalation at diagnosis. Circulating tumour cells (CTCs) found in the lymphovasculature are thought to have the propensity to metastasize at distant sites. CTCs are extremely rare events (typically a few CTCs per 10 6-9 blood cells), therefore very sensitive methodologies are required to isolate these cells [2]. Enrichment strategies aim to increase the CTC concentration from blood by several log units for easier detection by downstream methodologies. Biological approaches for CTC enrichment can include positive selection using anti-epithelial marker

ASSAY and Drug Development Technologies, 2016

Historically, two-dimensional (2D) cell culture has been the preferred method of producing diseas... more Historically, two-dimensional (2D) cell culture has been the preferred method of producing disease models in vitro. Recently, there has been a move away from 2D culture in favor of generating three-dimensional (3D) multicellular structures, which are thought to be more representative of the in vivo environment. This transition has brought with it an influx of technologies capable of producing these structures in various ways. However, it is becoming evident that many of these technologies do not perform well in automated in vitro drug discovery units. We believe that this is a result of their incompatibility with high-throughput screening (HTS). In this study, we review a number of technologies, which are currently available for producing in vitro 3D disease models. We assess their amenability with high-content screening and HTS and highlight our own work in attempting to address many of the practical problems that are hampering the successful deployment of 3D cell systems in mainstream research.

Journal of Solid Tumors, 2015

Dalton transactions (Cambridge, England : 2003), Jan 22, 2015

A first investigation into the application of a luminescent osmium(ii) bipyridine complex to live... more A first investigation into the application of a luminescent osmium(ii) bipyridine complex to live cell imaging is presented. Osmium(ii) (bis-2,2-bipyridyl)-2(4-carboxylphenyl) imidazo[4,5f][1,10]phenanthroline was prepared and conjugated to octaarginine, a cell penetrating peptide. The photophysics, cell uptake and cytotoxicity of this osmium complex conjugate were performed and compared with its ruthenium analogue. Cell uptake and distribution of both ruthenium and osmium conjugates were very similar with rapid transmembrane transport of the osmium probe (complete within approx. 20 min) and dispersion throughout the cytoplasm and organelles. The near-infrared (NIR) emission of the osmium complex (λmax 726 nm) coincides well with the biological optical window and this facilitated luminescent and luminescence lifetime imaging of the cell which was well resolved from cell autofluorescence. The large Stokes shift of the emission also permitted resonance Raman mapping of the dye within ...

British Journal of Cancer, 1998

Forty-two patients with hepatocellular carcinoma (HCC) were resected and their tumours were analy... more Forty-two patients with hepatocellular carcinoma (HCC) were resected and their tumours were analysed for p53 mutations by GCclamped denaturing gradient gel electrophoresis (DGGE), single-strand conformation polymorphism (SSCP) and gene sequencing. All the exons have been analysed in this study. Eight of 12 HCCs with cirrhosis due to viral hepatitis and the two patients with sarcomatoid changes displayed p53 mutations. In contrast, no mutation was observed in the fibrolamellar variant (n = 9), non-cirrhotics (n = 13) and alcoholic cirrhosis (n = 6). The mutations observed were in exons 5-8. Two mutations were observed in codons 136 and 213 as well as a T insertion between residues 156 and 157 (exon 5) and these are reported for the first time in HCC. Likewise, the silent mutation polymorphism in codon 213 was noticed in 3 of the 42 patients. Survival analysis of these patients after surgery showed the mean and median survival in patients with wild-type p53 to be 60 and 43 months respectively. In the group with p53 mutations, the mean and median survival was 15 and 12 months. The difference was statistically significant (P = 0.003).

ISCAS '98. Proceedings of the 1998 IEEE International Symposium on Circuits and Systems (Cat. No.98CH36187), 1998

A method is given to derive existence conditions for short-period oscillations which can occur in... more A method is given to derive existence conditions for short-period oscillations which can occur in the zero-input behaviour of a sigma-delta modulator structure incorporating a third order direct-form digital-filter. The approach is limited in practice to short periods and low-order digital filters because of the complexity of the symbolic and numerical analysis involved. However, short-period oscillations give rise to narrowband

Lecture Notes in Computer Science, 1996

There are still significant problems in the planning, design and management of public facilities ... more There are still significant problems in the planning, design and management of public facilities subject to dense pedestrian traffic. The automation of data collection and analysis of crowd behaviour is increasingly desirable in design of facilities and long-term site management using image processing techniques with existing closed-circuit television systems. We have investigated a number of techniques for crowd density estimation, movement estimation, incident detection and their relative merits using image processing. This paper presents techniques for background generation and calibration to enhance the previously-developed method of crowd density estimation using a reference image. An intensity region related to the average pixel intensity of each image in a sequence of crowd images is used to segment background pixels for generating a background image without pedestrians. The calibration approach, with which a previously-established relationship between image parameters and crowd density at one site can be used to estimate crowd density at various sites, involves calibration of the crowd image as opposed to calibration of the camera. Both techniques may be used in other surveillance systems such as vehicle monitoring.

Imaging Technology, Assay Development, and Data Analysis in Biology and Drug Discovery, 2014

European Journal of Biochemistry, 1981

Imaging Technology, Assay Development, and Data Analysis in Biology and Drug Discovery, 2014

Imaging Technology, Assay Development, and Data Analysis in Biology and Drug Discovery, 2014

European Journal of Cancer Supplements

Background: Tissue microarray analyses of human breast carcinomas have linked decreased annexin I... more Background: Tissue microarray analyses of human breast carcinomas have linked decreased annexin I (ANXA-1) expression with breast cancer progression [1]. ANXA-1, a calcium-and phospholipid-binding protein, has been implicated in A549 lung cancer cell growth and apoptosis [2]. We sought to establish the effect of ANXA-1 protein knockdown on cell cycle regulation in the estrogen receptor (ER)-positive MCF-7 cells and ERnegative MDA-MB-231 breast tumour cell lines. Materials & methods: MCF-7 and MDA-MB-231 cells were transfected with either ANXA-1 targeting stealth RNAi (siRNA) or its scrambled control sequence. After 24 hours, cells were stimuated with 5% FC. Cell lysates were obtained 24 and 48 hours post-transfection for analysis of ANXA-1 protein levels. Enumeration of viable cells was performed after a 48 hour incubation with FCS and cell cycle regulation was determined by flow cytometry of propidium iodide stained cells. Results: At 24 and 48 hours post-transfection, ANXA-1 protein levels were significantly decreased by 28% and 55% respectively in MCF-7 cells. The reduction of ANXA-1 in MDA-MB-231 cells at 48 hours after transfection was 35% (p < 0.05). ANXA-1 knockdown reduced basal and FCS-induced increases in MCF-7 cell number, accompanied, at 48 by significant reduction in G2/M phase cells. Neither basal nor FCS-induced increases in cell number were influenced by ANXA-1 siRNA in the ERnegative MDA-MB-231 cell line. Conclusions: These observations implicate ANXA-1 in the FCS-induced proliferation of MCF-7 cells. 1. Shen, D., et al., Decreased expression of annexin A1 is correlated with breast cancer development and progression as determined by a tissue microarray analysis.

Journal of Thoracic Oncology

Background: Bevacizumab (BEV), a humanized recombinant monoclonal antibody that targets vascular ... more Background: Bevacizumab (BEV), a humanized recombinant monoclonal antibody that targets vascular endothelial growth factor, is widely used in cancer treatment. BEV treatment is generally well-tolerated, however, patients who are treated with BEV have an increased risk of developing systemic adverse events, such as hypertension and proteinuria. We generally use angiotensin Iconverting enzyme inhibitors (ACEIs) and angiotensin II type-1 receptor blockers (ARBs) as a treatment of these adverse events. This group of drugs has been found to reduce proteinuria and prevent the development of glomeruloscrelosis. Additionally, since angiotensin II stimulates neovascularization, and thus act as a growth factor for cancer, previous studies have suggested that ACEIs and ARBs might decrease tumor growth and tumor-associated angiogenesis and inhibit metastasis. In this study, we aimed to investigate the correlation between proteinuria/hypertension and clinical outcome, and between ACEIs/ARBs use and clinical outcome in lung cancer patients treated with BEV-based chemotherapy. Methods: We retrospectively reviewed medical charts of patients at Kobe Red Cross Hospital and Kurashiki Central Hospital between November 2009 and February 2014. All patients were treated with BEV in combination with standard chemotherapy, such as pemetrexed plus platinum doublet, as first or second-line treatment. Cox regression analysis was performed to identify factors associated with antitumor response, overall survival (OS), and progression free survival (PFS). Results: Among 122 patients, almost 30 patients were excluded because of diabetes mellitus and use of .BEV in late lines. Ninety-nine patients were included in the analysis set. Median treated BEV cycles were 7 cycles. Median OS and median PFS were 13.0 months and 6.2 months, respectively. During the study, proteinuria was found in 56 patients (57%). Hypertension was found in 81 patients (82%). Thirty-nine patients (39%) were treated with ACEIs/ ARBs, 26 patients with other drugs. Univariate analysis showed that younger age, BEV cycles, and ACEIs/ARBs use were significantly associated with longer PFS, not OS. Younger age and BEV cycles were associated with PFS (P ¼ .030 and < 0.001, respectively) on multivariate analysis, however, proteinuria, hypertension, and ACEIs/ARBs use were not. There was however a trend for the correlation between ACEIs/ ARBs use and anti-tumor response (p ¼ 0.082). Conclusion: Our results suggest that BEV-related proteinuria and hypertension were not prognostic markers for lung cancer patients treated with BEV-based chemotherapy. Further accumulation of patients is needed to assess the potential effect of ACEIs/ARBs on anti-tumor effect and PFS.

Journal of Thoracic Oncology

Background: Stathmin 1 is a cytosolic phosphoprotein that plays a crucial role in the control of ... more Background: Stathmin 1 is a cytosolic phosphoprotein that plays a crucial role in the control of cellular division and proliferation by regulating microtubule dynamics. In addition, Stathmin1 is associated with tumor growth and progression. Our study aimed to determine differences in overall (OS) and disease free survival (DFS) in patients with non-small lung cancer (NSCLC) stratified by STMN1 tumor expression.

Nature Reviews Drug Discovery, 2016

The common and persistent failures to translate promising preclinical drug candidates into clinic... more The common and persistent failures to translate promising preclinical drug candidates into clinical success highlight the limited effectiveness of disease models currently used in drug discovery. An apparent reluctance to explore and adopt alternative cell-and tissuebased model systems, coupled with a detachment from clinical practice during assay validation, contributes to ineffective translational research. To help address these issues and stimulate debate, here we propose a set of principles to facilitate the definition and development of disease-relevant assays, and we discuss new opportunities for exploiting the latest advances in cell-based assay technologies in drug discovery, including induced pluripotent stem cells, 3D co-culture and organ-on-a-chip systems, complemented by advances in single-cell imaging and gene editing technologies. Funding to support precompetitive, multidisciplinary collaboration to develop novel preclinical models and cell-based screening technologies could have a key role in improving their clinical relevance, and ultimately increase clinical success rates [Au: edits to shorten OK?].

Oncotarget, 2014

Head and neck cancers are the seventh most common cancer globally. Approximately 50% of the patie... more Head and neck cancers are the seventh most common cancer globally. Approximately 50% of the patients die within 5 years. Current diagnostic methods include clinical assessment, imaging and tissue biopsy [1]. When metastases are clinically evident, treatment is palliative in nature only. There are currently no methods to predict which patients with a higher disease burden will develop metastases. The ability to do so would potentially lend itself to treatment escalation at diagnosis. Circulating tumour cells (CTCs) found in the lymphovasculature are thought to have the propensity to metastasize at distant sites. CTCs are extremely rare events (typically a few CTCs per 10 6-9 blood cells), therefore very sensitive methodologies are required to isolate these cells [2]. Enrichment strategies aim to increase the CTC concentration from blood by several log units for easier detection by downstream methodologies. Biological approaches for CTC enrichment can include positive selection using anti-epithelial marker

ASSAY and Drug Development Technologies, 2016

Historically, two-dimensional (2D) cell culture has been the preferred method of producing diseas... more Historically, two-dimensional (2D) cell culture has been the preferred method of producing disease models in vitro. Recently, there has been a move away from 2D culture in favor of generating three-dimensional (3D) multicellular structures, which are thought to be more representative of the in vivo environment. This transition has brought with it an influx of technologies capable of producing these structures in various ways. However, it is becoming evident that many of these technologies do not perform well in automated in vitro drug discovery units. We believe that this is a result of their incompatibility with high-throughput screening (HTS). In this study, we review a number of technologies, which are currently available for producing in vitro 3D disease models. We assess their amenability with high-content screening and HTS and highlight our own work in attempting to address many of the practical problems that are hampering the successful deployment of 3D cell systems in mainstream research.

Journal of Solid Tumors, 2015

Dalton transactions (Cambridge, England : 2003), Jan 22, 2015

A first investigation into the application of a luminescent osmium(ii) bipyridine complex to live... more A first investigation into the application of a luminescent osmium(ii) bipyridine complex to live cell imaging is presented. Osmium(ii) (bis-2,2-bipyridyl)-2(4-carboxylphenyl) imidazo[4,5f][1,10]phenanthroline was prepared and conjugated to octaarginine, a cell penetrating peptide. The photophysics, cell uptake and cytotoxicity of this osmium complex conjugate were performed and compared with its ruthenium analogue. Cell uptake and distribution of both ruthenium and osmium conjugates were very similar with rapid transmembrane transport of the osmium probe (complete within approx. 20 min) and dispersion throughout the cytoplasm and organelles. The near-infrared (NIR) emission of the osmium complex (λmax 726 nm) coincides well with the biological optical window and this facilitated luminescent and luminescence lifetime imaging of the cell which was well resolved from cell autofluorescence. The large Stokes shift of the emission also permitted resonance Raman mapping of the dye within ...

British Journal of Cancer, 1998

Forty-two patients with hepatocellular carcinoma (HCC) were resected and their tumours were analy... more Forty-two patients with hepatocellular carcinoma (HCC) were resected and their tumours were analysed for p53 mutations by GCclamped denaturing gradient gel electrophoresis (DGGE), single-strand conformation polymorphism (SSCP) and gene sequencing. All the exons have been analysed in this study. Eight of 12 HCCs with cirrhosis due to viral hepatitis and the two patients with sarcomatoid changes displayed p53 mutations. In contrast, no mutation was observed in the fibrolamellar variant (n = 9), non-cirrhotics (n = 13) and alcoholic cirrhosis (n = 6). The mutations observed were in exons 5-8. Two mutations were observed in codons 136 and 213 as well as a T insertion between residues 156 and 157 (exon 5) and these are reported for the first time in HCC. Likewise, the silent mutation polymorphism in codon 213 was noticed in 3 of the 42 patients. Survival analysis of these patients after surgery showed the mean and median survival in patients with wild-type p53 to be 60 and 43 months respectively. In the group with p53 mutations, the mean and median survival was 15 and 12 months. The difference was statistically significant (P = 0.003).

ISCAS '98. Proceedings of the 1998 IEEE International Symposium on Circuits and Systems (Cat. No.98CH36187), 1998

A method is given to derive existence conditions for short-period oscillations which can occur in... more A method is given to derive existence conditions for short-period oscillations which can occur in the zero-input behaviour of a sigma-delta modulator structure incorporating a third order direct-form digital-filter. The approach is limited in practice to short periods and low-order digital filters because of the complexity of the symbolic and numerical analysis involved. However, short-period oscillations give rise to narrowband

Lecture Notes in Computer Science, 1996

There are still significant problems in the planning, design and management of public facilities ... more There are still significant problems in the planning, design and management of public facilities subject to dense pedestrian traffic. The automation of data collection and analysis of crowd behaviour is increasingly desirable in design of facilities and long-term site management using image processing techniques with existing closed-circuit television systems. We have investigated a number of techniques for crowd density estimation, movement estimation, incident detection and their relative merits using image processing. This paper presents techniques for background generation and calibration to enhance the previously-developed method of crowd density estimation using a reference image. An intensity region related to the average pixel intensity of each image in a sequence of crowd images is used to segment background pixels for generating a background image without pedestrians. The calibration approach, with which a previously-established relationship between image parameters and crowd density at one site can be used to estimate crowd density at various sites, involves calibration of the crowd image as opposed to calibration of the camera. Both techniques may be used in other surveillance systems such as vehicle monitoring.

Imaging Technology, Assay Development, and Data Analysis in Biology and Drug Discovery, 2014

European Journal of Biochemistry, 1981

Imaging Technology, Assay Development, and Data Analysis in Biology and Drug Discovery, 2014

Imaging Technology, Assay Development, and Data Analysis in Biology and Drug Discovery, 2014