The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
All manuscripts are subject to rapid peer review. Those of high quality (not previously published and not under consideration for publication in another journal) will be published without delay.
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
All manuscripts are subject to rapid peer review. Those of high quality (not previously published and not under consideration for publication in another journal) will be published without delay.
In-vitro biological activities of the free new H4L ( indole-7-thiocarbohydrazone) ligand and its Ni(II), Pd(II) , Pt(II),
Cu(II), Ag(I), Zn(II) and Cd(II) complexes are screened against two cancerous cell lines, that revealed significant
activity only for [Cu2Cl2(H4L)2(PPh3)2] after 72 h treatment by the highest tested concentrations. The Copper(I)
complex was characterized by X-ray Crystallography and the NMR spectra, whereas it has been confirmed to have
momentous cytotoxicity against ovarian, breast cancerous cell lines (Caov-3, MCF-7). The apoptosis-inducing
properties of the Cu(I) complex have been investigated through fluorescence microscopy visualization, DNA
fragmentation analysis and propidium iodide flow cytometry.
BIOMIMESYS® Adipose tissue represents a new generation of mimetic Hydroscaffold™ for 3D culture of adipocyte and adipocyte-like cells. Available in a ready-to-use format, it enables the culture of adipocytes and adipocyte-like cells under physiological conditions that are representative of the microenvironment found in adipose tissue. The highly porous nature of the scaffold allows the rapid uptake of nutrients, oxygen, etc. into the cells to create a reproducible study model for all downstream analyses used with 3D adipocyte cultures.
Targeting PIM kinase to overcome drug resistance in NSCLC - Dr Kathy GatelyHannahMcCarthy31
Dr Kathy Gately is a Clinical Scientist at St James's Hospital and Clinical Senior Lecturer at Trinity College Dublin. Dr Gately's research interests are Drug Resistance Mechanisms in Non-Small Cell Lung Cancer, Liquid Biopsy and Organoid models.
NAD-Glycohydrolase Depletes Intracellular NAD+ and Inhibits Acidification of Autophagosomes to Enhance Multiplication of Group A Streptococcus in Endothelial Cells
Extraction of β-galactosidase and β-glucosidase from the seeds of Tamarindus ...Innspub Net
The enzymes β–galactosidase and β–glucosidase were extracted from the tamarind seeds using different buffers at different pH. Highest activity was obtained with 10 mM sodium acetate buffer, pH 5.6 and 10 mM tris buffer, pH 7.4. The effect of NaCl and Triton X–100 at different concentrations on the extraction of the enzymes indicated 10 mM sodium acetate buffer, pH 5.6 containing 1 M NaCl as a better extractant of the enzyme. The enzyme assay was carried out using p–nitrophenyl–β–D–galactoside and p–nitrophenyl–β–D–glucoside as substrates. Highest enzyme activities were observed on 6th and 24th day of germination. The protein content gradually decreased upto 5th day of germination and suddenly increased on 6th day. However, on subsequent days of germination, the protein content greatly decreased upto 11th day. During the latter period of germination (18th day onwards) the content remained almost constant. The kinetic parameters varied for both β–galactosidase and β–glucosidase. The activity of β–galactosidase was show to have an optimal operating condition at pH 5.5 and a temperature of 500C. The thermostability of the enzyme was in the range of 400C – 700C with the pH stability in the range of 5.0 – 7.0. The Km and Vmax values for pNPGal were determined as 66μM and 2.27nmolesmin-1. In contrast the activity of β–glucosidase was shown to have an optimal operating condition at pH 5.0 and a temperature of 300C. The thermostability of the enzyme was in the range of 270C – 350C with the pH stability in the range of 4.0 – 7.0. The Km and Vmax values for pNPGlu were determined as 121μM and 5.26nmolesmin-1. The presented study is a preliminary work carried out for the standardization of protocols. The purification and characterization of β–galactosidase and β–glucosidase is under progress.
IOMIMESYS® Oncology is a new generation of mimetic Hydroscaffolds™ for 3D cell culture. Available in a ready-to-use format, it enables the culture of cells under physiological conditions that are representative of the microenvironment found in whole tissues. The highly porous structure of BIOMIMESYS® allows cells to diffuse into the 3D matrix, where they can fix and begin to develop. The structure and conformation of cells cultured in BIOMIMESYS®, such as cancer cells that spontaneously form spheroids, strongly resemble the structures formed in vivo. Also, the functionality of the BIOMIMESYS® matrix is proved by the expression of genes and proteins at levels that are again more similar to those found in vivo. Cells cultured in BIOMIMESYS® give results that are more physiological than those grown in 2D culture and are closer to those obtained in vivo.
HepG2 cell model for genotoxicity and steatosis assessmentHCS Pharma
Early detection of toxic events induced by drug cantidats is mandatory in order to avoid late attrition in the process of R&D. Here we present two assays that can be done with the HepG2 human hepatoma cell line: genotoxicity assay (DNA double strand break) and steatosis.
Simplifying 3d cell culture generation for high content screening with BIOMIM...HCS Pharma
Growing interest in phenotypic screening, together with evidence that the drug response of cells grown in three-dimensional (3D) structures more closely resembles in vivo activity, has made high throughput, 3D fluorescence imaging an attractive screening option for drug discovery. However, creating 3D spheroids compatible with high content screening can be a difficult and expensive process.
BIOMIMESYS® is a range of patented hyaluronic acid scaffolds for 3D cell culture. They are made of RGDS- and galactosaminegrafted hyaluronic acid, using an adipic acid dihydrazide crosslinker and extracellular matrix proteins (eg. type I, IV or VI collagen) to accurately mimic the in vivo extracellular environment. BIOMIMESYS® is suitable for automated testing thanks to the uniform thickness of the scaffold – around 650 μm – with an average porosity of 100 to 200 μm
This application note describes a straightforward workflow for 3D cell seeding and spheroid formation using HCS Pharma’s BIOMIMESYS® plates in combination with INTEGRA’s VIAFLO 96/384 pipetting system. This process ensures rapid, controllable and reproducible 3D cell cultures, providing researchers with a highly efficient method to produce physiologically-relevant cellular models in a high throughput format
In-vitro biological activities of the free new H4L ( indole-7-thiocarbohydrazone) ligand and its Ni(II), Pd(II) , Pt(II),
Cu(II), Ag(I), Zn(II) and Cd(II) complexes are screened against two cancerous cell lines, that revealed significant
activity only for [Cu2Cl2(H4L)2(PPh3)2] after 72 h treatment by the highest tested concentrations. The Copper(I)
complex was characterized by X-ray Crystallography and the NMR spectra, whereas it has been confirmed to have
momentous cytotoxicity against ovarian, breast cancerous cell lines (Caov-3, MCF-7). The apoptosis-inducing
properties of the Cu(I) complex have been investigated through fluorescence microscopy visualization, DNA
fragmentation analysis and propidium iodide flow cytometry.
BIOMIMESYS® Adipose tissue represents a new generation of mimetic Hydroscaffold™ for 3D culture of adipocyte and adipocyte-like cells. Available in a ready-to-use format, it enables the culture of adipocytes and adipocyte-like cells under physiological conditions that are representative of the microenvironment found in adipose tissue. The highly porous nature of the scaffold allows the rapid uptake of nutrients, oxygen, etc. into the cells to create a reproducible study model for all downstream analyses used with 3D adipocyte cultures.
Targeting PIM kinase to overcome drug resistance in NSCLC - Dr Kathy GatelyHannahMcCarthy31
Dr Kathy Gately is a Clinical Scientist at St James's Hospital and Clinical Senior Lecturer at Trinity College Dublin. Dr Gately's research interests are Drug Resistance Mechanisms in Non-Small Cell Lung Cancer, Liquid Biopsy and Organoid models.
NAD-Glycohydrolase Depletes Intracellular NAD+ and Inhibits Acidification of Autophagosomes to Enhance Multiplication of Group A Streptococcus in Endothelial Cells
Extraction of β-galactosidase and β-glucosidase from the seeds of Tamarindus ...Innspub Net
The enzymes β–galactosidase and β–glucosidase were extracted from the tamarind seeds using different buffers at different pH. Highest activity was obtained with 10 mM sodium acetate buffer, pH 5.6 and 10 mM tris buffer, pH 7.4. The effect of NaCl and Triton X–100 at different concentrations on the extraction of the enzymes indicated 10 mM sodium acetate buffer, pH 5.6 containing 1 M NaCl as a better extractant of the enzyme. The enzyme assay was carried out using p–nitrophenyl–β–D–galactoside and p–nitrophenyl–β–D–glucoside as substrates. Highest enzyme activities were observed on 6th and 24th day of germination. The protein content gradually decreased upto 5th day of germination and suddenly increased on 6th day. However, on subsequent days of germination, the protein content greatly decreased upto 11th day. During the latter period of germination (18th day onwards) the content remained almost constant. The kinetic parameters varied for both β–galactosidase and β–glucosidase. The activity of β–galactosidase was show to have an optimal operating condition at pH 5.5 and a temperature of 500C. The thermostability of the enzyme was in the range of 400C – 700C with the pH stability in the range of 5.0 – 7.0. The Km and Vmax values for pNPGal were determined as 66μM and 2.27nmolesmin-1. In contrast the activity of β–glucosidase was shown to have an optimal operating condition at pH 5.0 and a temperature of 300C. The thermostability of the enzyme was in the range of 270C – 350C with the pH stability in the range of 4.0 – 7.0. The Km and Vmax values for pNPGlu were determined as 121μM and 5.26nmolesmin-1. The presented study is a preliminary work carried out for the standardization of protocols. The purification and characterization of β–galactosidase and β–glucosidase is under progress.
IOMIMESYS® Oncology is a new generation of mimetic Hydroscaffolds™ for 3D cell culture. Available in a ready-to-use format, it enables the culture of cells under physiological conditions that are representative of the microenvironment found in whole tissues. The highly porous structure of BIOMIMESYS® allows cells to diffuse into the 3D matrix, where they can fix and begin to develop. The structure and conformation of cells cultured in BIOMIMESYS®, such as cancer cells that spontaneously form spheroids, strongly resemble the structures formed in vivo. Also, the functionality of the BIOMIMESYS® matrix is proved by the expression of genes and proteins at levels that are again more similar to those found in vivo. Cells cultured in BIOMIMESYS® give results that are more physiological than those grown in 2D culture and are closer to those obtained in vivo.
HepG2 cell model for genotoxicity and steatosis assessmentHCS Pharma
Early detection of toxic events induced by drug cantidats is mandatory in order to avoid late attrition in the process of R&D. Here we present two assays that can be done with the HepG2 human hepatoma cell line: genotoxicity assay (DNA double strand break) and steatosis.
Simplifying 3d cell culture generation for high content screening with BIOMIM...HCS Pharma
Growing interest in phenotypic screening, together with evidence that the drug response of cells grown in three-dimensional (3D) structures more closely resembles in vivo activity, has made high throughput, 3D fluorescence imaging an attractive screening option for drug discovery. However, creating 3D spheroids compatible with high content screening can be a difficult and expensive process.
BIOMIMESYS® is a range of patented hyaluronic acid scaffolds for 3D cell culture. They are made of RGDS- and galactosaminegrafted hyaluronic acid, using an adipic acid dihydrazide crosslinker and extracellular matrix proteins (eg. type I, IV or VI collagen) to accurately mimic the in vivo extracellular environment. BIOMIMESYS® is suitable for automated testing thanks to the uniform thickness of the scaffold – around 650 μm – with an average porosity of 100 to 200 μm
This application note describes a straightforward workflow for 3D cell seeding and spheroid formation using HCS Pharma’s BIOMIMESYS® plates in combination with INTEGRA’s VIAFLO 96/384 pipetting system. This process ensures rapid, controllable and reproducible 3D cell cultures, providing researchers with a highly efficient method to produce physiologically-relevant cellular models in a high throughput format
Digital Catapult Centre Brighton: Retail Innovation and the Store of The Futu...wired_sussex
The Digital Catapult Centre Brighton is about collaborative R&D and enabling breakthrough innovations.
At this event we introduced Store of the Future - a physical space in the Netherlands that hosts innovative retail products and services.
http://storeofthefuture.nl/
Watch for an introductory Store of the Future talk by it’s Retail Research Director John Terra.
Follow us on Twitter to get involved in the conversation
https://twitter.com/Digicatbrighton
If you want to learn more about the Digital Catapult Centre Brighton, check out the website here: http://www.digitalcatapultcentre.org....
Don't forget to follow us on Twitter to join in the conversation and to receive live updates: https://twitter.com/Digicatbrighton
We also have a Linkedin group, please join to engage and share ideas with other participants interested in the Digital Catapult Centre Brighton: https://www.linkedin.com/grp/home?gid...
If you would like to be added to our Slack group, got any questions about any of our events or have just any questions in general, feel free to drop us an email at any time.
Rosalie@wiredsussex.com
Tech Beyond The Screen: Real-Time Place-Based Digital Marketing Henry Bennett...wired_sussex
At The Digital Catapult Centre Brighton event, Tech Beyond The Screen: Real-Time Place-Based Digital Marketing Henry Bennett talked about his company Your Welcome.
The Electrophoretic Profile Myofibrillar Proteins Extracted From Camel Muscle...IJEAB
Changes in electrophoretic profiles of myofibrillar protein (MFP) in the Longissimus thoracis (LD) of young camels (2 to 4 years), preserved by refrigeration has been treated or not by lactic acid solution 4% or citric acid 1%, were followed during the post-mortem time at the following times: 1, 2, 4, 6, 8, 10, 12, 24 and 48 hours. The cold preservation for 48 hours has not shown any particular distinctions in the protein profiles of this muscle. Changes related to the type of treatment were recorded during the storage time. Proteolysis of the myofibrillar fraction was earlier in this muscle in the case of treatment with one of two solutions of organic acids used, particularly in the case of using lactic acid. Indeed, these changes have affected at the first hour after slaughter the proteolysis of the myofibrillar proteins. Fragments of low molecular weight (42, 36, 33, 26, 23, 18, 16, 14 and 13 kDa) have been identified. The electrophoretic analysis showed that during refrigeration, LD treated with a solution of lactic acid is more sensitive to disruption phenomena and muscle protein proteolysis that lots of this muscle that even in the case of preservation by refrigeration only or by refrigeration
Objective: To investigate the protective effect of lo- sartan, an angiotensin II type 1 receptor blocker with antioxidative effect on intestinal ischemia-reperfusion (I/R) injury in rats, against inflammation and apoptotic development.
Study Design: Forty male Wistar albino rats with a mean weight of 200–250 g each were divided into 4 groups: (1) Sham operation (laparotomy only, sham surgical preparation including isolation of the superior mesenteric artery [SMA] without occlusion), (2) Ischemia model with SMA closure for 2 hours, (3) I/R group (2 hours of ischemia followed by 3-hour reperfusion (SMA occlusion for 120 minutes followed by 240 minutes reperfusion), and (4) Losartan group (2 hours of ischemia, 40 mg/kg losartan was administered to the animals; losartan was dissolved in 1 mL distilled water and administered intraperitoneally after 2 hours of ischemia). Malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) levels were examined in jejunum tissue.
Results: Losartan treatment reduced the I/R-induced increase in MDA levels in the gut. Statistically, while SOD, CAT, and GSH activities decreased significantly in the I/R group, they increased in the I/R+Losartan group. Villus loss and increase in inflammation after ischemia persisted after reperfusion. Losartan treatment played a role in the reduction of inflammation and apoptosis and in the regulation of TNF-α and caspase-9 activity.
Conclusion: It has been thought that losartan in I/R may reduce mucosal damage and cell apoptosis in the direction of inflammation and may stabilize caspase-9 activity by inhibiting TNF-α stimulus.
Keywords: caspase-9, ischemia, ischemia/reperfusion, rat, reperfusion injury, TNF-α, tumor necrosis factor-alpha
TNF-alpha and LPA promote synergistic expression of COX-2 in human colonic my...Enrique Moreno Gonzalez
Enhanced EGF receptor (EGFR) signaling is a hallmark of many human cancers, though the role of enhanced EGFR signaling within the surrounding tumor stroma has not been well studied. The myofibroblast is an important stromal cell that demonstrates enhanced EGFR expression in the setting of inflammation, though the functional relevance is not known. We
recently reported that TNF-α and the G protein-coupled receptor (GPCR) agonist lysophosphatidic acid (LPA) lead to synergistic cyclo-oxygenase-2 (COX-2) expression, an enzyme strongly associated with the development of colitis-associated cancer. Here, we investigate whether EGFR signaling plays a role in the synergistic COX-2 expression induced by LPA and TNF-α.
BIOMIMESYS®Liver, a 3D cell culture model for maintaining and promoting hepat...HCS Pharma
BIOMIMESYS® range are hyaluronan based hydroscaffold developed to overcome the 2D flat culture limitations by recreating an in vivo-like physiology within the in vitro environment.
BIOMIMESYS®Liver hydroscaffold is made of RGDS and galactosamine-grafted Hyaluronic acid, Adipic acid dihydrazide crosslinker and extracellular matrix (ECM) proteins (collagen type I and collagen type IV) to mimic liver ECM composition.
BIOMIMESYS® Liver, a 3D cell culture model for maintaining and promoting hep...HCS Pharma
BIOMIMESYS® range are hyaluronan based hydroscaffold developed to overcome the 2D flat culture limitations by recreating an in vivo-like physiology within the in vitro environment.
BIOMIMESYS®Liver scaffold is made of RGDS and galactosamine-grafted Hyaluronic acid, Adipic acid dihydrazide crosslinker and extracellular matrix (ECM) proteins (collagen type I and collagen type IV) to mimic liver-ECM composition.
Normal tissues and tumors arise from a population of cells termed stem cells. In vivo experiments have provided evidence of the presence of stem cells throughout the mouse mammary gland. Premalignant mammary outgrowths that faithfully recapitulate the mammary epithelial cell lineage upon transplantation contain cells with tumor-forming potential. Cell sorting techniques have identified putative mouse mammary stem cell surface markers and human breast cancer stem cell surface markers. These markers do not identify only stem cells but in fact distinguish a mixed population of cells containing stem cell activity. Previous studies have demonstrated that clones arising from single cells in vitro can be categorized into three types based on the clone morphology. Here, we report the characterization, both in vitro and in vivo, of clonogenic cells from a non-tumorigenic mammary epithelial population and those from an erbB2-induced mammary tumor. We found that clones arising from normal mammary cells expressed different patterns of stem and developmental marker between the clone types and compared to the expression patterns observed on clones that developed from tumorigenic mammary cells.
ABSTRACT- The anticancer drug arsenic trioxide is effective for acute promyelocytic leukemia. But the clinical trials are
restricted due to its potential side effects. Since the major part of arsenic metabolism and detoxification occurs in liver,
this organ faces the major threat. The hepatic side effects include fatty liver, fibrosis, and inflammation and hepatocyte
degeneration. Our study aimed to evaluate the protective potential of the fatty acid, docosahexaenoic acid, against adversities
of arsenic trioxide in an in vitro model, the Chang liver cells. Two preliminary dose standardization assays, cell
viability and lactate dehydrogenase release assays, were employed. The assays were performed as Pre-treatment,
Co-treatment and Post treatment experiments for a period of 24 hours. Arsenic trioxide at various doses (2.5, 5, 7.5, 10,
12.5 and 15 μM) showed a significant (p≤0.05) dose dependant reduction in cell viability along with a dose dependant
enhancement of lactate dehydrogenase release. However when the cells were treated with a combination of docosahexaenoic
acid at varying concentrations (50, 75, 100, 125 and 150 μM), the above mentioned conditions were found to be
reversed in Pre-treatment and Co-treatment experiments, but not in Post treatment. The most effective combination was
found to be 10 μM arsenic trioxide with 100 μM of docosahexaenoic acid in both Pre-treatment and Co- treatment studies.
Thus the preliminary assays of our study showed that docosahexaenoic acid administration as Pre-treatment or
Co-treatment can aid in reducing arsenic trioxide induced hepatotoxicity. Further studies are required to elucidate the mechanisms
behind the protective effects.
Key Words– Arsenic trioxide, hepatotoxicity, docosahexaenoic acid, cell damage
Similar to Inhibition mmp 2 and mmp-9 of different molecule weigh function by membrane system from abalone haliotis discus hannai in the ht1080 cells (20)
Sudheer Mechineni, Head of Application Frameworks, Standard Chartered Bank
Discover how Standard Chartered Bank harnessed the power of Neo4j to transform complex data access challenges into a dynamic, scalable graph database solution. This keynote will cover their journey from initial adoption to deploying a fully automated, enterprise-grade causal cluster, highlighting key strategies for modelling organisational changes and ensuring robust disaster recovery. Learn how these innovations have not only enhanced Standard Chartered Bank’s data infrastructure but also positioned them as pioneers in the banking sector’s adoption of graph technology.
Generative AI Deep Dive: Advancing from Proof of Concept to ProductionAggregage
Join Maher Hanafi, VP of Engineering at Betterworks, in this new session where he'll share a practical framework to transform Gen AI prototypes into impactful products! He'll delve into the complexities of data collection and management, model selection and optimization, and ensuring security, scalability, and responsible use.
A tale of scale & speed: How the US Navy is enabling software delivery from l...sonjaschweigert1
Rapid and secure feature delivery is a goal across every application team and every branch of the DoD. The Navy’s DevSecOps platform, Party Barge, has achieved:
- Reduction in onboarding time from 5 weeks to 1 day
- Improved developer experience and productivity through actionable findings and reduction of false positives
- Maintenance of superior security standards and inherent policy enforcement with Authorization to Operate (ATO)
Development teams can ship efficiently and ensure applications are cyber ready for Navy Authorizing Officials (AOs). In this webinar, Sigma Defense and Anchore will give attendees a look behind the scenes and demo secure pipeline automation and security artifacts that speed up application ATO and time to production.
We will cover:
- How to remove silos in DevSecOps
- How to build efficient development pipeline roles and component templates
- How to deliver security artifacts that matter for ATO’s (SBOMs, vulnerability reports, and policy evidence)
- How to streamline operations with automated policy checks on container images
UiPath Test Automation using UiPath Test Suite series, part 4DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 4. In this session, we will cover Test Manager overview along with SAP heatmap.
The UiPath Test Manager overview with SAP heatmap webinar offers a concise yet comprehensive exploration of the role of a Test Manager within SAP environments, coupled with the utilization of heatmaps for effective testing strategies.
Participants will gain insights into the responsibilities, challenges, and best practices associated with test management in SAP projects. Additionally, the webinar delves into the significance of heatmaps as a visual aid for identifying testing priorities, areas of risk, and resource allocation within SAP landscapes. Through this session, attendees can expect to enhance their understanding of test management principles while learning practical approaches to optimize testing processes in SAP environments using heatmap visualization techniques
What will you get from this session?
1. Insights into SAP testing best practices
2. Heatmap utilization for testing
3. Optimization of testing processes
4. Demo
Topics covered:
Execution from the test manager
Orchestrator execution result
Defect reporting
SAP heatmap example with demo
Speaker:
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
GraphSummit Singapore | The Art of the Possible with Graph - Q2 2024Neo4j
Neha Bajwa, Vice President of Product Marketing, Neo4j
Join us as we explore breakthrough innovations enabled by interconnected data and AI. Discover firsthand how organizations use relationships in data to uncover contextual insights and solve our most pressing challenges – from optimizing supply chains, detecting fraud, and improving customer experiences to accelerating drug discoveries.
GraphSummit Singapore | The Future of Agility: Supercharging Digital Transfor...Neo4j
Leonard Jayamohan, Partner & Generative AI Lead, Deloitte
This keynote will reveal how Deloitte leverages Neo4j’s graph power for groundbreaking digital twin solutions, achieving a staggering 100x performance boost. Discover the essential role knowledge graphs play in successful generative AI implementations. Plus, get an exclusive look at an innovative Neo4j + Generative AI solution Deloitte is developing in-house.
GridMate - End to end testing is a critical piece to ensure quality and avoid...ThomasParaiso2
End to end testing is a critical piece to ensure quality and avoid regressions. In this session, we share our journey building an E2E testing pipeline for GridMate components (LWC and Aura) using Cypress, JSForce, FakerJS…
LF Energy Webinar: Electrical Grid Modelling and Simulation Through PowSyBl -...DanBrown980551
Do you want to learn how to model and simulate an electrical network from scratch in under an hour?
Then welcome to this PowSyBl workshop, hosted by Rte, the French Transmission System Operator (TSO)!
During the webinar, you will discover the PowSyBl ecosystem as well as handle and study an electrical network through an interactive Python notebook.
PowSyBl is an open source project hosted by LF Energy, which offers a comprehensive set of features for electrical grid modelling and simulation. Among other advanced features, PowSyBl provides:
- A fully editable and extendable library for grid component modelling;
- Visualization tools to display your network;
- Grid simulation tools, such as power flows, security analyses (with or without remedial actions) and sensitivity analyses;
The framework is mostly written in Java, with a Python binding so that Python developers can access PowSyBl functionalities as well.
What you will learn during the webinar:
- For beginners: discover PowSyBl's functionalities through a quick general presentation and the notebook, without needing any expert coding skills;
- For advanced developers: master the skills to efficiently apply PowSyBl functionalities to your real-world scenarios.
PHP Frameworks: I want to break free (IPC Berlin 2024)Ralf Eggert
In this presentation, we examine the challenges and limitations of relying too heavily on PHP frameworks in web development. We discuss the history of PHP and its frameworks to understand how this dependence has evolved. The focus will be on providing concrete tips and strategies to reduce reliance on these frameworks, based on real-world examples and practical considerations. The goal is to equip developers with the skills and knowledge to create more flexible and future-proof web applications. We'll explore the importance of maintaining autonomy in a rapidly changing tech landscape and how to make informed decisions in PHP development.
This talk is aimed at encouraging a more independent approach to using PHP frameworks, moving towards a more flexible and future-proof approach to PHP development.
Why You Should Replace Windows 11 with Nitrux Linux 3.5.0 for enhanced perfor...SOFTTECHHUB
The choice of an operating system plays a pivotal role in shaping our computing experience. For decades, Microsoft's Windows has dominated the market, offering a familiar and widely adopted platform for personal and professional use. However, as technological advancements continue to push the boundaries of innovation, alternative operating systems have emerged, challenging the status quo and offering users a fresh perspective on computing.
One such alternative that has garnered significant attention and acclaim is Nitrux Linux 3.5.0, a sleek, powerful, and user-friendly Linux distribution that promises to redefine the way we interact with our devices. With its focus on performance, security, and customization, Nitrux Linux presents a compelling case for those seeking to break free from the constraints of proprietary software and embrace the freedom and flexibility of open-source computing.
Epistemic Interaction - tuning interfaces to provide information for AI supportAlan Dix
Paper presented at SYNERGY workshop at AVI 2024, Genoa, Italy. 3rd June 2024
https://alandix.com/academic/papers/synergy2024-epistemic/
As machine learning integrates deeper into human-computer interactions, the concept of epistemic interaction emerges, aiming to refine these interactions to enhance system adaptability. This approach encourages minor, intentional adjustments in user behaviour to enrich the data available for system learning. This paper introduces epistemic interaction within the context of human-system communication, illustrating how deliberate interaction design can improve system understanding and adaptation. Through concrete examples, we demonstrate the potential of epistemic interaction to significantly advance human-computer interaction by leveraging intuitive human communication strategies to inform system design and functionality, offering a novel pathway for enriching user-system engagements.
State of ICS and IoT Cyber Threat Landscape Report 2024 previewPrayukth K V
The IoT and OT threat landscape report has been prepared by the Threat Research Team at Sectrio using data from Sectrio, cyber threat intelligence farming facilities spread across over 85 cities around the world. In addition, Sectrio also runs AI-based advanced threat and payload engagement facilities that serve as sinks to attract and engage sophisticated threat actors, and newer malware including new variants and latent threats that are at an earlier stage of development.
The latest edition of the OT/ICS and IoT security Threat Landscape Report 2024 also covers:
State of global ICS asset and network exposure
Sectoral targets and attacks as well as the cost of ransom
Global APT activity, AI usage, actor and tactic profiles, and implications
Rise in volumes of AI-powered cyberattacks
Major cyber events in 2024
Malware and malicious payload trends
Cyberattack types and targets
Vulnerability exploit attempts on CVEs
Attacks on counties – USA
Expansion of bot farms – how, where, and why
In-depth analysis of the cyber threat landscape across North America, South America, Europe, APAC, and the Middle East
Why are attacks on smart factories rising?
Cyber risk predictions
Axis of attacks – Europe
Systemic attacks in the Middle East
Download the full report from here:
https://sectrio.com/resources/ot-threat-landscape-reports/sectrio-releases-ot-ics-and-iot-security-threat-landscape-report-2024/
Threats to mobile devices are more prevalent and increasing in scope and complexity. Users of mobile devices desire to take full advantage of the features
available on those devices, but many of the features provide convenience and capability but sacrifice security. This best practices guide outlines steps the users can take to better protect personal devices and information.
2. Fish Aquat Sci 15(2), 137-143, 2012
Materials
loproteinases has also been called type IV collagenases, because of their ability to cleave type IV collagen (Salo et al.,
1985). One of them, MMP-2 (gelatinase-A, 72 kDa type IV
collagenase), was originally purified from a highly progressing metastatic murine tumor (Wilhelm et al., 1989). MMP-2
binds to type I collagen through the fibronectin domain, which
stabilizes it against autolysis, thereby controlling its activity
(Ellerbroek et al., 2001). MMP-2 expression is dependent on
extracellular matrix metalloproteinase inducer, growth factors, cytokines, and hormones. Pro-MMP-2 activation needs
contributions from MT1-MMP and TIMP metallopeptidase
inhibitor 2. A low level has been linked to a favorable prognosis in patients with hormone receptor-negative tumors, usually
associated with poor prognosis. As a zymogen that requires
proteolytic activation for catalytic activity, MMP-2 has been
implicated in invasion and metastasis in many cancer model
systems, including human breast cancer (Jezierska and Motyl, 2009; Fredrich and Illing, 2010). MMP-9 (92 kDa type IV
collagenase, gelatinase B) is produced in human macrophages
and polymorphonuclear leukocytes. It has also been localized
in endothelial cells and synovial fibroblasts in rheumatoid arthritis synovium. MMP-9 is expressed by osteoclasts in human normal bone tissues, indicating a role in bone remodelling. Intact mature human odontoblasts also express MMP-9,
and it has been detected in human dental caries lesions and saliva. However, MMP-9 is not expressed by human gingival fibroblasts. Like MMP-2, MMP-9 may exist in the ECM bound
to type I collagen, gelatine, or laminin (Bolcato-Bellemin et
al., 2000).
Degradation of denatured collagen I by MMP-2 and MMP9 is readily demonstrable in biological materials. One technique, known as gelatin zymography, identifies gelatinolytic
activity in biological samples using sodium dodecyl sulfate
(SDS)-polyacrylamide gels impregnated with gelatine. The
human fibrosarcoma cell line HT1080 has been used extensively in studies of ECM proteins involved in attachment, invasion, and metastasis. Additionally, HT1080 cells are used
because they produce both MMP-2 and MMP-9 enzymes
(Brooks and Schumacher, 2001).
The abalone Haliotis discus hannai, which is maricultured
on the southwestern coast (Wando area) of Korea, is a marine univalve gastropod mollusc, that inhabits temperate and
tropical waters in both hemispheres of the globe. Despite its
economic importance, few nutraceutical and pharmaceutical
studies of abalone have been reported.
In the present study, to investigate the bioactive potential of
abalone intestine, abalone intestine G I digests (AIGIDs) were
separated into various molecular weight (MW) fractions by
ultrafiltration. We focuses on AIGID II and III mediated inhibition of both MMP-2 and MMP-9 and cancer cell migration
in human fibrosarcoma HT1080 cells.
Live adult abalones were collected from Wando Island, Korea. Intestinal organs (guts) were separated from the washed
abalone and lyophilized. Dulbecco’s modified Eagle’s medium
(DMEM), trypsin-ethylenediaminetetraacetic acid (EDTA),
penicillin/streptomycin, and fetal bovine serum (FBS) were
obtained from Gibco BRL, Life Technologies (Grand Island,
NY, USA). HT1080 cells were obtained from the American
Type Culture Collection (Manassas, VA, USA). Primary and
secondary antibodies, including MMP-2 (sc-13595), MMP-9
(sc-10737), NF-κB p65 (sc-8008), NF-κB p50 (sc-166588),
β-actin (sc-130656), goat anti-rabbit IgG-horseradish peroxidase (HRP) (sc-2004) and goat anti-mouse IgG1-HRP (sc2060), were purchased from Santa Cruz Biotechnology-Inc
(Santa Cruz, CA, USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), gelatin (type A) and phorbol 12-myristate 13-acetate (PMA) were from Sigma Chemical Co. (St. Louis, MO, USA). Other chemicals and reagents
used were of analytical grade.
Preparation of AIGIDs using ultrafiltration (UF)
membrane bioreactor
The method of Kapsokefalou and Miller (1991) was used
to prepare AIGIDs (AIGID and AIGID I-III). Abalone intestine solution (1 L) was adjusted to pH 2.2 for gastric digestion
using 1 M HCl and 10 M NaOH. Pepsin (EC 3.4.23.1) was
added at an enzyme:substrate ratio of 1:100 (w/w), then incubated at 37°C on a shaker for 2 h. The pH was adjusted to 6.5
to match the conditions of small intestinal digestion. Trypsin
(EC 3.4.21.4) and α-chymotrypsin (EC 3.4.21.1) were added
at an enzyme to substrate ratio of 1/100 (w/w), then incubated
at 37°C for 2.5 h. AIGIDs were centrifuged (10,000 g, 15 min,
at 4°C), and the supernatant was lyophilized to obtain AIGID
powder. AIGID (100 g) was solublized in 2 L of water and
fractionated using UF membranes with a 100 kDa MW cut-off
(MWCO). The filtrate was then fractionated further through a
10 kDa MWCO filter. All samples (AIGID and AIGID I-III)
were dialyzed through a 1 kDa membrane and lyophilized.
Cell culture
Human fibrosarcoma HT1080 cells were cultured in DMEM
containing 10% FBS and antibiotics as monolayers in 10 cm
culture dishes at 37°C in a humidified 5% CO2 atmosphere.
MTT assay
Cytotoxic effects of the AIGIDs on cultured cells were
measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. To determine the cytocompatible effects of AIGIDs, HT1080 cells were seeded
in 96-well plates at a 1×104 /well and incubated with various
Materials and Methods
http://dx.doi.org/10.5657/FAS.2012.0137
138
3. Nguyen et al. (2012) Inhibitory effects of abalone intestine digests on MMPs expression in fibrosarcoma cells
AIGID, AIGID I, AIGID II, and AIGID III concentrations. After a 24 h incubation, MTT reagent (50 µL) was added to each
well and incubation was continued for another 4 h. Finally,
DMSO was added to solubilize the formazan salt formed and
the absorbance at 540 nm was determined using a microplate
reader. Cell viability relative to the non-treated group was calculated. Data are expressed as means of at least three independent experiments and P < 0.05 were considered to indicate
statistical significance.
Gelatin zymography
The gelatinase activities of MMP-2 and MMP-9 were determined by gelatin zymography (Ta et al., 2006; Kong et al.,
2010). Cells were cultured on 24-well plates in serum-free
DMEM medium. HT1080 cells were treated with AIGID,
AIGID I, AIGID II, and AIGID III for 1 h and then stimulated
with 10 ng/mL PMA for 36 h. The conditioned media were
collected and centrifuged (3000 g, 10 min). Concentrated
medium was electrophoresed under non-reducing conditions
through 10% sodium dodecyl sulfate (SDS)-polyacrylamide
gels impregnated with 0.15% of gelatin. After electrophoresis,
SDS was removed from the gel by washing in 2.5% Triton
X-100 solution for 1.5 h and incubated in developing buffer
(50 mM Tris-HCl, pH 7.5, 200 mM NaCl, 5 mM CaCl2.2H2O,
0.02% Brif-35) at 37°C. The gels were stained with 1% Coomassie blue R-250 in 45% methanol and 10% glacial acetic
acid. After 30 min, the gels were destained in the same solution with no Coomassie blue dye. The digested area appeared
clear on a blue background, indicating the presence of a gelatinase.
Fig. 1. Effects of abalone intestine G I digests (AIGIDs; AIGID, AIGID I,
AIGID II and AIGID III) on viability of HT1080 cells. Cells were treated with
different concentrations (10-400 μg/mL) of AIGIDs and cell viability was
determined by MTT assay after 24 h. Deta are give as means of values ±
SD. from three independent experiments.
dilution) in blocking agent at 4°C overnight. After washing
with TBS-T, the membrane was incubated with secondary antibody (1:5,000 dilution) for 2 h at room temperature. Bands
were developed by enhanced chemiluminescence and visualized with LAS-4000 imaging system (Fujifilm, Tokyo, Japan).
Basal levels of MMP-2 and MMP-9 protein expression were
normalized by assessing the level of β-actin protein.
Amino acid composition
Freeze-dired AIGID II and AIGID III (20 mg) was hydrolyzed with 6 M HCl at 110°C for 24 h in vacuum. Amino acids
derived with phenylisothiocyanate were identified and quantified using an automatic amino acid analyzer (Biochrom 20;
Pharmacia Biotech, Cambridge, UK).
In vitro wound migration assay
HT1080 cells were seeded in six-well plates. Cells were
pretreated with mitomycin C (25 μg/mL) for 30 min before
an injury line was made with a 2 mm wide tip on cells that
were plated in culture dishes at 80% confluence. Cells were
then washed with phosphate buffered saline. The cells were
allowed to migrate in serum-free medium in the presence of
various AIGID II and AIGID III concentrations for 24 h. Results were observed by microscopy.
Results
Cytocompatible effects of AIGIDs in HT1080 cells
The effects of AIGID and AIGID I-III on proliferation of
HT1080 cells were evaluated using an MTT assay. No AIGID
had any cytotoxic effect on HT1080 cells up to 400 μg/mL
(Fig. 1).
Western blot analysis
AIGIDs inhibit MMP-2 and MMP-9 activity in
HT1080 cells
Cultured cells were harvested and then lysed with RIPA
buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1% Triton
X-100, 1% sodium deoxycholate, 2 mM EDTA, and 0.1%
SDS). Protein concentration was determined by the BCA
method. Proteins were separated by 10% SDS-polyacrylamide gel electrophoresis and transferred to a nitrocellulose
membrane. Membranes were blocked with 5% skim milk in
TBS-T buffer (20 mM Tris-HCl, pH 7.6, 136 mM NaCl, 0.1%
Tween-20) and then incubated with primary antibody (1:500
The effects of AIGIDs of differing MWs on MMP-2 and
MMP-9 were evaluated. Cells treated with AIGID, AIGID I,
AIGID II, or AIGID III at 200 μg/mL were stimulated with
PMA. Conditioned media was used to assess MMP-2 and -9
activities. AIGID II and AIGID III showed marked inhibitory
effects on MMP-2 and MMP-9 in HT1080 cells compared with
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4. Fish Aquat Sci 15(2), 137-143, 2012
In vitro effects of AIGID II and III on migration of
HT1080 cells
the PMA treatment group (Fig. 2). However, neither AIGID
nor AIGID I significantly inhibited MMP-2 or MMP-9. Thus,
AIGID II and III were used in subsequent experiments.
A gelatin zymography assay was performed to assess the
dose-dependent inhibitory activities of AIGID II and III on
MMP-2 and MMP-9 mRNA expression in HT1080 cells.
MMP-2 and MMP-9 expression was suppresed by AIGID II
and III in a dose-dependent manner (Fig.3). AIGID III showed
greater inhibition of MMP-2 and MMP-9 expression than did
AIGID II at 200 μg/mL.
Cell migration is required for cancer cell invasion through
the basement membrane. HT 1080 cells were treated with
0-200 μg/mL AIGID II and III. Both AIGID II and III inhibited the migration of HT1080 cells at 200 μg/mL (Fig. 4).
EffectS of AIGID II and III on MMP-2 and MMP-9
protein expression and NF-κB activity in HT1080
cells
AIGID II and III mediated inhibition of MMP-2 and MMP9 protein levels were analyzed by Western blotting. AIGID II
and III (10, 100, 200 μg/mL) suppressed MMP-2 and MMP-9
protein levels in a concentration-dependent manner, compared
with PMA treatment (Fig. 5). Inhibition of MMP-2 expression
by both AIGID II and III (200 μg/mL) was more noticeable.
Furthermore, Western blots were used to assess the downregulation of protein expression and to determine the possible
mechanisms for the effects of AIGID II and III on the expression of activator NF-κB. Western blotting was conducted to
confirm down-regulation of p50 and p65 protein expression;
both are components of NF-кB. AIGID II and III treatment
resulted in decreased p65 protein level; this was dose-dependent (10, 100, 200 μg/mL) (Fig. 6). Additionally, AIGID III
showed a greater inhibition of p65 expression than AIGID II at
200 μg/mL. However, no significant inhibition of p50 expression was detected at the concentrations tested. These results
demonstrate that AIGID II and III down-regulated MMP-2
and MMP-9 expression through transcriptional down-regulation of NF-κB.
Fig. 2. Gelatin zymography for the determination of matrix metallopro-
teinase (MMP)-2 and -9 activities in abalone intestine G I digest (AIGID)
treated HT1080 cells. HT1080 cells treated with 200 μg/mL of AIGID, AIGID
I, AIGID II and AIGID III for 1 h were stimulated by phorbol 12-myristate
13-acetate (PMA; 10 ng/mL) for 36 h. Gelatinolytic activities of MMP-2 and
MMP-9 in conditioned media were detected by electrophoresis of soluble
protein on a gelatine containing 10% polyacrylamide gel.
A
Amino acid composition
B
The amino acid compositions of AIGID II and III are shown
in Table 1. AIGID II and III were rich in acidic amino acids
(glutamic acid, aspartic acid), proline (Pro), alanine (Ala), histidine (His), and lysine (Lys), which constituted 63.12% of the
total amono acid residues in AIGID II and 56.58 % of AIGID
III. The amino acid composition of a protein is associated with
its biological activities; indeed, Glu, Ala, and His have been
reported to be important for the inhibition of MMP-2 and -9
(Emara and Cheung, 2006).
Discussion
Fig. 3. Effects of abalone intestine G I digest (AIGID) II and AIGID III on
matrix metalloproteinase (MMP)-2 and MMP-9 activities by gelatin zymography. (A) Gelatinase activities of MMP-2 and MMP-9 in conditioned
media were detected by electrophoresis of soluble protein on a gelatin
containing 10% polyacrylamide gel. (B) Areas and relative intensities of
gelatin-digested bands by MMP-2 and MMP-9 were quantified by densitometry and expressed as relative MMP-9 activity compared to that of
phorbol 12-myristate 13-acetate (PMA)-alone treated cells.
http://dx.doi.org/10.5657/FAS.2012.0137
In this study, we demonstrated that AIGID II and III markedly decreased MMP expression, cell migration, motility and
invasiveness (Fig. 4). Tumor cell invasion of the ECM is an
important step in tumor metastasis, and involves the attachment of tumor cells to ECM (Cavallaro and Christofori, 2001;
Zhang et al., 2009). Tumor cell migration and invasion are
140
5. Nguyen et al. (2012) Inhibitory effects of abalone intestine digests on MMPs expression in fibrosarcoma cells
dependent on gelatinase activity. MMP-2 and MMP-9 are
important for tumor metastasis. MMPs are highly expressed
in human cancers, and a direct relationship between cancer
progression and MMP expression and activity has been established by many studies (Rajapakse et al., 2007). The present
study confirmed the multi-inhibitory effect of AIGID II and III
on MMP-2 and MMP-9 expression using an artificial substrate
and gelatin zymography. In PMA-treated HT1080 cells, AIGID II and III suppressed the expression and secretion of MMP2 and MMP-9 by down regulating of MMP-9 transcriptional
activation in a dose-dependent manner (Fig. 5). The MMP-2
and MMP-9 promoters both contains NF-κB sites.
Fig. 4. Effects of abalone intestine G I digest (AIGID) II and AIGID III on migration of HT1080 cells. After pretreatment of cells with mitomycin C (25 μg/mL)
for 30 min, injury line was made on the confluent monolayer of cells. HT1080 cells were treated with AIGID II and AIGID III for 24 h. Cell motility was examined with light microscope at indicated time points.
Table 1. Amino acid composition of abalone intestine gastrointestinal
A
digests (AIGIDs)
AIGIDs (mg/100 g)
Amino acid
AIGID II
AIGID III
Asp
6.95
5.95
Thr-
4.07
4.66
Ser
1.75
2.38
Glu
B
13.57
16.17
Gly
2.97
5.68
Ala
11.35
10.55
Cys
5.45
4.4
Val
0.5
2.25
3.54
Leu
4.83
6.99
Tyr
2.83
2.66
Phe
3.28
3.79
His
9.78
4.97
Lys
8.7
4.9
Trp-
2.94
1.76
Arg
2 and MMP-9 protein expression in HT1080 cell after treatment various
concentrations of abalone intestine G I digest (AIGID) II and AIGID III for
1 h and stimulated with phorbol 12-myristate 13-acetate (PMA; 10 ng/
mL). Expression of β-actin protein was the control for normalization of
MMP-2 and MMP-9 protein. (B) Areas and intensities of protein bands
were determined by densitometry and expressed as a percentage MMP
expression compared to protein levels of PMA-alone treated cells. Values
represent means ± SEM from two independent experiments.
4.58
1.85
Iie
Fig. 5. (A) Western blot analysis of matrix metalloproteinase (MMP)-
3.2
Met
1.64
2.5
Pro
Total
12.77
100
14.04
100
AIGID II (100-10 kDa) and AIGID III (10-5 kDa) by the in vitro GI digestion
model (SHGD).
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6. Fish Aquat Sci 15(2), 137-143, 2012
and are necessary for gene induction. NF-κB regulates critical processes in cancers, including invasion, metastasis, apoptosis, angiogenesis, growth, and proliferation (Hayden and
Ghosh, 2004; Mendis et al., 2009). NF-κB triggers MMP-2
and MMP-9 gene transcription in response to chemical stimuli, such as PMA, in HT1080 cells (Kim and Kim, 2010). The
promoter of the MMP-9 gene contains several putative binding sites for transcription factors that regulate gene expression, an AP-1 binding consensus site at -79 bases upstream
from the starter site. Further upstream there is a cluster of regulatory elements, including AP-1, AP-2. and NF-κB binding
sites. Additionally, in some inflammatory conditions, MMP-9
transcription can be regulated by NF-κB via different signalling pathways (Hayden and Ghosh, 2004). Thus, we evaluated the ability of AIGID to down-regulate NF-κB pathways.
AIGID suppression was clearly dose-dependent. However, it
did not suppress PMA-induced translocation of p50, also a
component of NF-κB (Fig. 6). MMP-2 and MMP-9 activities
were reduced in the presence of some amino acids including
cysteine and histidine, cysteine had the strongest inhibitory
effect on both MMP- 2 and -9 expression (Emara and Cheung,
2006). Both the differential inhibition of MMP-2 and MMP-9
expression by AIGID we report here and the molecular structure of some amino acids (cysteine and histidine), are consistent with this.
In conclusion, the AIGID II and III fractions of AIGIDs inhibited the PMA-induced invasion and migration of HT1080
cells. Furthermore, AIGID II and III inhibited MMP-2 and
MMP-9 expression through down-regulation of the transcription factor NF-κB. Therefore, both AIGID and its active components may be useful as anti-invasive agents in therapeutic
strategies against fibrosarcoma metastasis.
A
B
Fig. 6. Western blot analysis of nuclear transcription factor kappa B (NFκB; p65 and p50) protein expressions in HT1080 cells treated with abalone
intestine G I digest (AIGID) II and AIGID III with different concentrations.
(A) Equal amounts of protein in the cell lysates were electrophoresed and
p50 and p65 protein levels in cytosol were determined using specific antibodies. Expression of β-actin protein was the control the equal amounts
of protein used for electrophoresis. (B) Areas and intensities of protein
bands were determined by densitometry and expressed as a percentage
NF-κB expression compared to protein levels of phorbol 12-myristate
13-acetate (PMA)-alone treated cells. Values represent means ± SEM from
two independent experiments.
Acknowledgments
The human fibrosarcoma HT1080 cell line produces large
quantities of MMP-9, whereas normal human fibroblasts have
been reported to secrete little or none (Moll et al., 1990). Several studies have shown that PMA can also induce expression
of MMP-2 and MMP-9, another important MMP involved in
cancer progression. However, the expression level of MMP-2
in HT1080 cells is lower than that of MMP-9 (Kong et al.,
2008; Kong et al., 2010). However, under our experimental
conditions, in HT1080 cells with DMEM plus 10% FBS,
MMP-2 expression was lower than that of MMP-9 following
PMA treatment and that in the no-treatment group (Fig. 2).
PMA enhanced MMP-9 expression by about 40% compared
with that in non-stimulated cells, over time (Fig. 3). This elevated MMP-9 expression was inhibited in a dose-dependent
manner by all AIGID II and III concentrations tested.
Furthermore, the NF-κB family of transcription factors has
been shown to be present in most cell types, and specific NFκB binding sites, termed κB sites, have been identified in the
promoters of a large number of inducible genes. p50 and p65
are two NF-κB subunits that contain transactivation domains
http://dx.doi.org/10.5657/FAS.2012.0137
This Study was supported by Technology Development
Program for Fisheries, Ministry for Food, Agriculture, Forestry and Fisheries, and a grant from the Korean Ministry
of Knowledge and Economy through the Program for the
Regional Innovation System (RIS). It was also supported by
Wando-County Program for R&D Services, Wando-gun, Jeonnam, Republic of Korea.
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