Abstract: Adipose-derived stem cells (ADSCs), were isolated from discarded human fat tissue, obtained from csection with our recently modified methods, in Stem Cell & Regenerative Medicine Lab, VSBT. Here we develop
two methods to isolate Adipose derived mesenchymal stem cells with enzyme digestion and use of
phosphatidylcholine and deoxycholate. Surface protein expression was analyzed by flow cytometry to
characterize the cell phenotype. The multi-lineage potential of ADSCs was testified by differentiating cells with
adipogenic inducer. ADSCs can be cultured in vitro for up to one month without passage. Also, the flow
cytometry analysis showed that ADSCs expressed high levels of stem cell related surface marker CD105.
ADSCs have strong proliferation ability, maintain their phenotypes, and have stronger multi-differentiation
potential. The molecular basis of ADSC differentiation was studied using bioinformatics tools with an aim to
identify the key proteins involved in differentiation, such that they could be used as potential targets for drug
development for the treatment of obesity. The key proteins involved were found to be PPARG and C/EBPα. The
structures of the proteins were retrieved from MMDB (Molecular Modelling Database) and PDB (Protein Data
Bank) respectively. Key Words: Adipose-derived stem cells, Mesenchymal stem cells, Enzyme digestion, Phosphatidylcholine, Deoxycholate, PPARG, C/EBPα, etc.
Poster – Development and automation of 3D innovative hiPSC-based liver organo...HCS Pharma
We previously showed that human pluripotent stem cells (hiPSCs) provide a suitable model to study metabolic diseases upon hepatocyte-like cell (HLC) differentiation. In particular, HLCs have been used to model cholesterol metabolism regulation, by mimicking the main disease features in vitro. Human iPSCs can be generated from urine samples of patients with a well-described phenotype and carrying specific genotypes. This non-invasive approach allowed the study of LDLR- and PCSK9-mediated autosomal dominant hypercholesterolemia (ADH) as well as PCSK9-mediated familial hypobetalipoproteinemia (FHBL). While the direct link between hiPSCs and patients, as well as the abundance of HLCs provide promising advantages of such strategy, it is impaired mainly by the neonatal characteristic of HLCs as well as the difficulty to perform high throughput studies for pharmacological investigations.
Induced Pluripotent Stem Cells and Somatic Cardiac Regeneration— An Explorato...Robert Chen
In the exploratory analysis, the functional pathway networks of nucleostemin and iPS interact via TP53 (tumor protein P53) and Fgf (fibroblast growth factor), which could be further investigated to provide clues for the future research of postnatal cardiac regeneration.
International Journal of Stem Cell Research and Transplantation (IJST) ISSN:2328-3548, is a free, Open Access, Peer-reviewed, exclusive online journal covering areas of Stem cell research, translational work and Clinical studies in the specialty of Stem Cells and Transplantation including allied specialties relevant to the core subject, which is dedicated in publishing high quality manuscripts.
International Journal of Stem Cell Research and Transplantation (IJST) is a peer-reviewed journal, and is dedicated to providing information with respect to the latest advancements that are being upgraded in our everyday life with respect to the application of Stem cells.
International Journal of Stem Cell Research and Transplantation (IJST) is an international, Open Access, peer-reviewed journal, which mainly focuses, on the advancements made in the field of cell biology, specifically in the field of Stem Cells.
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
Study of Various Histogram Equalization TechniquesIOSR Journals
Abstract: Histogram equalization (HE) works well on single channel images for contrast enhancement. However, the technique used is ineffective on multiple channel images. So, it is not suitable for consumer electronic products, where preserving the original brightness is necessary in order not to introduce unnecessary visual deterioration. Bi-histogram equalization (BHE) has been developed and it is analyzed mathematically.BHE separates the input image’s histogram into two, based on its mean before equalizing them independently so that it can preserve the original brightness up to certain extends. Recursive Mean-Separate Histogram Equalization (RMSHE) is another technique to provide better and scalable brightness preservation for gray scale and color images. While the separation is done only once in BHE, RMSHE performs the separation recursively based on their respective mean. It is analyzed mathematically that the output images mean brightness will converge to the input images mean brightness as the number of recursive mean separation increases. The recursive nature of RMSHE also allows scalable brightness preservation, which is very useful in consumer electronics. Finally a comparative study was made to analyze all the above methods using gray scale and color images. Keywords: Bi-histogram equalization, histogram equalization, scalable brightness preservation, recursive mean-separate
Poster – Development and automation of 3D innovative hiPSC-based liver organo...HCS Pharma
We previously showed that human pluripotent stem cells (hiPSCs) provide a suitable model to study metabolic diseases upon hepatocyte-like cell (HLC) differentiation. In particular, HLCs have been used to model cholesterol metabolism regulation, by mimicking the main disease features in vitro. Human iPSCs can be generated from urine samples of patients with a well-described phenotype and carrying specific genotypes. This non-invasive approach allowed the study of LDLR- and PCSK9-mediated autosomal dominant hypercholesterolemia (ADH) as well as PCSK9-mediated familial hypobetalipoproteinemia (FHBL). While the direct link between hiPSCs and patients, as well as the abundance of HLCs provide promising advantages of such strategy, it is impaired mainly by the neonatal characteristic of HLCs as well as the difficulty to perform high throughput studies for pharmacological investigations.
Induced Pluripotent Stem Cells and Somatic Cardiac Regeneration— An Explorato...Robert Chen
In the exploratory analysis, the functional pathway networks of nucleostemin and iPS interact via TP53 (tumor protein P53) and Fgf (fibroblast growth factor), which could be further investigated to provide clues for the future research of postnatal cardiac regeneration.
International Journal of Stem Cell Research and Transplantation (IJST) ISSN:2328-3548, is a free, Open Access, Peer-reviewed, exclusive online journal covering areas of Stem cell research, translational work and Clinical studies in the specialty of Stem Cells and Transplantation including allied specialties relevant to the core subject, which is dedicated in publishing high quality manuscripts.
International Journal of Stem Cell Research and Transplantation (IJST) is a peer-reviewed journal, and is dedicated to providing information with respect to the latest advancements that are being upgraded in our everyday life with respect to the application of Stem cells.
International Journal of Stem Cell Research and Transplantation (IJST) is an international, Open Access, peer-reviewed journal, which mainly focuses, on the advancements made in the field of cell biology, specifically in the field of Stem Cells.
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
Study of Various Histogram Equalization TechniquesIOSR Journals
Abstract: Histogram equalization (HE) works well on single channel images for contrast enhancement. However, the technique used is ineffective on multiple channel images. So, it is not suitable for consumer electronic products, where preserving the original brightness is necessary in order not to introduce unnecessary visual deterioration. Bi-histogram equalization (BHE) has been developed and it is analyzed mathematically.BHE separates the input image’s histogram into two, based on its mean before equalizing them independently so that it can preserve the original brightness up to certain extends. Recursive Mean-Separate Histogram Equalization (RMSHE) is another technique to provide better and scalable brightness preservation for gray scale and color images. While the separation is done only once in BHE, RMSHE performs the separation recursively based on their respective mean. It is analyzed mathematically that the output images mean brightness will converge to the input images mean brightness as the number of recursive mean separation increases. The recursive nature of RMSHE also allows scalable brightness preservation, which is very useful in consumer electronics. Finally a comparative study was made to analyze all the above methods using gray scale and color images. Keywords: Bi-histogram equalization, histogram equalization, scalable brightness preservation, recursive mean-separate
The Prevalence of Alcohol Consumption among Commercial Drivers in Uyo Local G...IOSR Journals
Abstract: The purpose of the study was to assess the prevalence of alcohol consumption among commercial
drivers in Uyo metropolis. Five research questions and three null hypothesis design was adopted, and the
instrument for the study was mainly interview schedules.
Due to the transitory nature of drivers in Uyo motor parks, convenient sampling was used to draw 160 drivers
who use Uyo motor parks.
The descriptive statistics percentage was used to answer the research questions, while chi – square statistics
was used to test the hypotheses at 0.05 level of significance. All the drivers interviewed drink alcohol for several
reasons. The sale of alcohol in the park and its environs has significant (P< 0.05) influence on their use. There
is no statistically significant difference (P>0.05) in the perceived influence of the use of alcohol on health with
respect to years of experience and age of drivers. The study was concluded with appropriate recommendation to
help the situation.
Key words: alcohol, drivers, prevalence and Uyo
Comparative Assessment of Two Thermodynamic Cycles of an aero-derivative Mari...IOSR Journals
Abstract: This paper explores the gas turbine potentials that are fully enhanced by the use of intercooling and
thermal recuperation as an engineering option available in the design of gas turbines and offered for marine
applications. It examines the off-design performance of two different cycle designs of a 25MW aero-derivative
engine by modelling and simulating each of them to operate under conditions other than those of their design
point. The simple cycle model consists of a single-spool dual shaft layout while the advanced model is
represented by an intercooled-recuperated cycle that runs on a dual-spool and is driven through a three shaft
configuration. In each case, the output shaft is coupled to a power turbine through which the propulsion power
may be transmitted to the propeller of the vessel to operate in a virtual marine environment. An off-design
performance simulation of both engines has been conducted in order to investigate and compare the effect of
ambient temperature variation during their part-load operation and particularly when subjected to a variety of
marine operating conditions. The study assesses the techno-economic impact of the complex design of the
advanced cycle over its simple cycle counterpart and demonstrates its potential for improved operating cost
through reduced fuel consumption as a significant step in the current drive for establishing the marine gas
turbine engine as a viable alternative to traditional prime movers in the ship propulsion industry.
Wear Analysis of Polytetrafluoroethylene (PTFE) and it’s Composites under Wet...IOSR Journals
In this paper, the effect of load, Velocity of sliding and sliding distance on friction and wear of
materials made of Polytetrafluoroethylele (PTFE) and PTFE composites under wet condition with filler
materials such as 25% bronze, 25% glass fiber and 25 % carbon have studied. The experimental work has
performed on pin-on-disc friction and wear test rig and analyzed with the help of Design Expert software. The
results of experiments are presented in tables and graphs which shows that the addition of bronze, glass and
carbon filler to the virgin PTFE decreases wear rate significantly and there is marginal increase in coefficient
of friction. The highest wear resistance was found for 25% carbon filled PTFE followed by 25% glass filled
PTFE, 25% bronze filled PTFE and virgin PTFE. Through this study, we can develop the best bearing material
for the various industrial applications which is available easily at the minimum cost.
Field Programmable Gate Array for Data Processing in Medical SystemsIOSR Journals
Abstract: Two- dimensional &Three–dimensional (3-D) image segmentation is on of the most demanding tasks in image processing. It has been proven that only the 14-neighborship of a rhombic dodecahedron can satisfy the aforementioned requirements. The 3-D-GSC process is executed in the following three phases ,coding phases,linking phases,splitting phases. An FPGA-based digital signal processing board optimized for applications needing large memory with high bandwidth has been developed and successfully used for the parallelization of a modern image segmentation algorithm for medical and industrial real-time applications.The Use of this 128-bit coprocessor board is not limited to image segmentation.We propose the perfectly parallelizable 3-D Gray-Value Structure Code (3-D-GSC) for image segmentation on a new FPGA custom machine. This 128-Bit FPGA coprocessing board features an up-to-date Virtex-II Pro architecture, two large independent DDR-SDRAM channels, two fast independent ZBT-SRAM channels, and PCI-X bus and CameraLink interfaces. Key words: Field Programmable Gate Array, Segme-ntation, Vogel bruch, Gray-valve structure code ,Homogeneous, Linking phase, Coding phase, Splitting phase, SDRAM.
Calcium Chloride Applications to Improve Fruit Quality on Bruised and Disease...IOSR Journals
The problems faced by producer canned of pineapple are the high of bruised which caused by the mechanical damage such as pressure, vibration during harvest, transport to the fruit processing and pathological damage caused by fruit diseases. The objective of research was to obtain the treatment time of CaCl2 applications and dosage of CaCl2 to improve the fruit quality of pineapple. This research used Split Plot Design and each treatment replicated 3 times. The main plot is time of CaCl2 applications that consists of three levels, thats are : 90 day after forcing (daf) (W90), 120 daf (W120) and twice time of CaCl2 applications on 90 and 120 daf (W90+120). The sub plot is dosage of CaCl2 that consists of three levels, thats are : 50 kg ha-1 (C50), 75 kg ha-1 (C75) and 100 kg ha-1 (C100). The results of research showed that the combined treatment twice time of CaCl2 applications on 90 and 120 day after forcing and dosage of CaCl2 100 kg ha-1 produces the calcium content on fruit is higher than the other combined treatments and produce the fruit texture, percentage of fruit diseases and percentage of bruised are lower than the other combined treatments
Implementation of XOR Based Pad Generation Mutual Authentication Protocol for...IOSR Journals
In RF link, without security the messages exchange between the two devices are monitoring by an
eavesdropper. So the exclusive-OR (XOR) based pad generation protocol is used to safely transfer the data to
the other point with necessary security and it maintaining confidentiality. This protocol produce the cover
coding pad to mask the access password before the datas are transmitted. A specially designed pad generation
will be implemented in digital domain to solve the insecurity problem in data communication RF link. This
protocol developed under regulation of ISO 18000 – 6 type C protocol also known as EPC C1G2 RFID
protocol. The linear feed back shift register (LFSR) generate the pseudo random binary sequence (PRBS) and it
is used as data source to the pad generation function. The Xilinx 13.x software is used for synthesize and
modelsim SE6.0 is used for simulating the result. The pad generation algorithm has been implemented in FPGA
Spartan 3 PQ208-4 board to verify the result
Comparative Study and Analysis of Image Inpainting TechniquesIOSR Journals
Abstract: Image inpainting is a technique to fill missing region or reconstruct damage area from an image.It
removes an undesirable object from an image in visually plausible way.For filling the part of image, it use
information from the neighboring area. In this dissertation work, we present a Examplar based method for
filling in the missing information in an image, which takes structure synthesis and texture sysnthesis together.
In exemplar based approach it used local information from an image to patch propagation.We have also
implement Nonlocal Mean approach for exemplar based image inpainting.In Nonlocal mean approach it find
multiple samples of best exemplar patches for patch propagation and weight their contribution according to
their similarity to the neighborhood under evaluation. We have further extended this algorithm by considering
collaborative filtering method to synthesize and propagate with multiple samples of best exemplar patches. We
have to preformed experiment on many images and found that our algorithm successfully inpaint the target
region.We have tested the accuracy of our algorithm by finding parameter like PSNR and compared PSNR
value for all three different approaches.
Keywords: Texture Synthesis, Structure Synthesis, Patch Propagation ,imageinpainting ,nonlocal approach,
collabrative filtering.
Blow Mould Tool Design and Manufacturing Process for 1litre Pet BottleIOSR Journals
the concepts of Blow molding is a process used to produce hollow objects from thermoplastic. The
basic blow molding process has two fundamental phases. First, a parson (or a perform) of hot plastic resin in a
somewhat tubular shape is created. Second, compressed air is used to expand the hot perform and press it
against mould cavities. The pressure is held until the plastic cools. Blow molding process is used for which has
thin wall sections.In this thesis, blow mould design is to be done for a bottle having 0.5mm thickness. This
thickness cannot be filled in pressure injection molding. So blow molding is considered for pet bottle design.
The mould is prepared by first modeling the part, extracting core & cavity and generating CNC program. Blow
mould tool design is done in Pro/Engineer according to HASCO standards. A prototype of the pet bottle using
blow mould design is also included.
Depth Estimation and Source Location of Magnetic Anomalies from a Basement Co...IOSR Journals
Source locations and depths to magnetic contacts were estimated from the total intensity magnetic
field of an area of 3,025.25 square kilometres on geographical latitude 7o00' N to N o ' 7 30 and longitude
E o ' 3 00 to E o ' 3 30 within Abeokuta area, using local wavenumber method. This study was carried out using
digitised airborne magnetic data of basement complex formation. Structural interpretation of the magnetic data
was achieved through applying advanced processing techniques that provide automatic delineation and depth
estimation of the magnetic structures. Local wavenumber method was used for locating and estimating depth to
magnetic contact. The magnetic contact depth ranges from 0.145km to 2.692km.
Accretion Profile of the Rosy Barb, Puntius Conchonius (Hamilton- Buchanan, 1...IOSR Journals
Present study was made on 50 freshwater wild Puntius conchonius (Hamilton - Buchanan) of
various sizes ranging from a total length of 5.2 cm to 7.6 cm and weighing 1.6 gm to 7.1 gm. They were
sampled from different lentic and lotic water bodies of Nagaland, to investigate the weight-length relationship.
Each fish was measured and weight was taken. Log transformed regressions were used to test the growth trend. It was observed that growth in weight is not proportional to the cube of its length. Coefficient of correlation values for both male and female as well as for mixed population were found to be highly significant.
Inventory Management System and Performance of Food and Beverages Companies i...IOSR Journals
Inventory management decisions are an integral aspect of organisations. Inventory postponement as
argued by Bucklin (1965) is where a firm deliberately delays the purchase and the physical possession of
inventory items until demand or usage requirements are known with certainty. This is an effective supply chain
strategy adopted by most manufacturing organisations by reducing the inventory, and in turn reducing the cost
of obsolete stock. This study explores the relationship between inventory management and control and
performance and Food and Beverages companies in Nigeria. Secondary data were obtained from annual
financial reports and accounts of Food and Beverages companies listed on the Nigerian Stock Exchange. The
data obtained were analyzed using simple and multiple regression models. The results show that there
significant relationship between inventory management and control and the performance of Food and
Beverages companies in Nigeria. The multiple regression correlation coefficient (R) =0.996, R2=0.990 and pvalue
=0;00<0.05 The results also show the relative importance of the inventory management decisions made
by the organisation, and the implications these decisions have on the consumer. The findings show that the three
key qualities that are essential in inventory management decisions for manufacturing organisation from the
perspective of the third party logistics provider are customer satisfaction, on time delivery and order fulfillment
Half-metallic-ferrimagnetic Sr2CrWO6 and Sr2FeReO6 materials for room tempera...IOSR Journals
Complex perovskite-like materials which include magnetic transition elements have relevance due to
the technological perspectives in the spintronics industry. In this work, we report the studies of the electronic
and magnetic characterizations of Sr2CrWO6 and Sr2FeReO6 as spintronics materials at room temperature by
using the linearized muffin-tin orbitals (LMTO) method through the atomic-sphere approximation (ASA) within
the local spin density approximation (LSDA). The interchange-correlation potential was included through the
LSDA+U technique. The band structure results at room-temperature predict half-metallic ferrimagnetic ground
state for Sr2CrWO6 and Sr2FeReO6 with total magnetic moment of 1.878 μB and 3.184 μB per formula unit,
respectively, agreement with the previous theoretical and experimental results.
Parametric sensitivity analysis of a mathematical model of facultative mutualismIOSR Journals
The complex dynamics of facultative mutualism is best described by a system of continuous non-linear first order ordinary differential equations. The methods of 1-norm, 2-norm, and infinity-norm will be used to quantify and differentiate the different forms of the sensitivity of model parameters. These contributions will be presented and discussed.
Reprogramming to pluripotency is possible from adult cells of different tissues and species through the ectopic expression of defined factors. The generated induced Pluripotent Stem Cells (iPSCs) are relevant for various purposes, including disease modeling, drug or toxicity screening and autologous cell therapy. Over the last few years, increased efforts are being made to improve the reprogramming techniques, the efficiency and quality of the generated iPSCs, as well as to identify the best cell source to be reprogrammed. Cells derived from fetal tissues, such as amniotic fluid, placenta and umbilical cord, offer distinct advantages in terms of reprogramming compared to adult somatic cells. Importantly, fetal cells are more primitive, easily achievable in sufficient numbers and are devoid of any ethical concern. They show great plasticity, high proliferation rate, low immunogenity and absence of teratoma formation. Therefore, they can be reprogrammed much faster and more efficiently than adult cells. Here, we provide a comprehensive overview of the advantages of reprogramming fetal sources in comparison to other commonly used cell types.
Meritxell Huch - Wellcome Trust/Cancer Research UK Gurdon Institute, Universi...Fundación Ramón Areces
El jueves 8 de febrero de 2018 se realizó en la Fundación Ramón Areces un Ciclo de Conferencias sobre células madre y organoides, en colaboración con Springer Nature.
Organs-on-chips (OoCs) are systems containing engineered or natural miniature tissues grown inside microfluidic chips. To better mimic human physiology, the chips are designed to control cell microenvironments and maintain tissue-specific functions. Combining advances in tissue engineering and microfabrication, OoCs have gained interest as a next-generation experimental platform to investigate human pathophysiology and the effect of therapeutics in the body. There are as many examples of OoCs as there are applications, making it difficult for new researchers to understand what makes one OoC more suited to an application than another.
SAP Sapphire 2024 - ASUG301 building better apps with SAP Fiori.pdfPeter Spielvogel
Building better applications for business users with SAP Fiori.
• What is SAP Fiori and why it matters to you
• How a better user experience drives measurable business benefits
• How to get started with SAP Fiori today
• How SAP Fiori elements accelerates application development
• How SAP Build Code includes SAP Fiori tools and other generative artificial intelligence capabilities
• How SAP Fiori paves the way for using AI in SAP apps
Pushing the limits of ePRTC: 100ns holdover for 100 daysAdtran
At WSTS 2024, Alon Stern explored the topic of parametric holdover and explained how recent research findings can be implemented in real-world PNT networks to achieve 100 nanoseconds of accuracy for up to 100 days.
In his public lecture, Christian Timmerer provides insights into the fascinating history of video streaming, starting from its humble beginnings before YouTube to the groundbreaking technologies that now dominate platforms like Netflix and ORF ON. Timmerer also presents provocative contributions of his own that have significantly influenced the industry. He concludes by looking at future challenges and invites the audience to join in a discussion.
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.
Accelerate your Kubernetes clusters with Varnish CachingThijs Feryn
A presentation about the usage and availability of Varnish on Kubernetes. This talk explores the capabilities of Varnish caching and shows how to use the Varnish Helm chart to deploy it to Kubernetes.
This presentation was delivered at K8SUG Singapore. See https://feryn.eu/presentations/accelerate-your-kubernetes-clusters-with-varnish-caching-k8sug-singapore-28-2024 for more details.
Le nuove frontiere dell'AI nell'RPA con UiPath Autopilot™UiPathCommunity
In questo evento online gratuito, organizzato dalla Community Italiana di UiPath, potrai esplorare le nuove funzionalità di Autopilot, il tool che integra l'Intelligenza Artificiale nei processi di sviluppo e utilizzo delle Automazioni.
📕 Vedremo insieme alcuni esempi dell'utilizzo di Autopilot in diversi tool della Suite UiPath:
Autopilot per Studio Web
Autopilot per Studio
Autopilot per Apps
Clipboard AI
GenAI applicata alla Document Understanding
👨🏫👨💻 Speakers:
Stefano Negro, UiPath MVPx3, RPA Tech Lead @ BSP Consultant
Flavio Martinelli, UiPath MVP 2023, Technical Account Manager @UiPath
Andrei Tasca, RPA Solutions Team Lead @NTT Data
GraphRAG is All You need? LLM & Knowledge GraphGuy Korland
Guy Korland, CEO and Co-founder of FalkorDB, will review two articles on the integration of language models with knowledge graphs.
1. Unifying Large Language Models and Knowledge Graphs: A Roadmap.
https://arxiv.org/abs/2306.08302
2. Microsoft Research's GraphRAG paper and a review paper on various uses of knowledge graphs:
https://www.microsoft.com/en-us/research/blog/graphrag-unlocking-llm-discovery-on-narrative-private-data/
GDG Cloud Southlake #33: Boule & Rebala: Effective AppSec in SDLC using Deplo...James Anderson
Effective Application Security in Software Delivery lifecycle using Deployment Firewall and DBOM
The modern software delivery process (or the CI/CD process) includes many tools, distributed teams, open-source code, and cloud platforms. Constant focus on speed to release software to market, along with the traditional slow and manual security checks has caused gaps in continuous security as an important piece in the software supply chain. Today organizations feel more susceptible to external and internal cyber threats due to the vast attack surface in their applications supply chain and the lack of end-to-end governance and risk management.
The software team must secure its software delivery process to avoid vulnerability and security breaches. This needs to be achieved with existing tool chains and without extensive rework of the delivery processes. This talk will present strategies and techniques for providing visibility into the true risk of the existing vulnerabilities, preventing the introduction of security issues in the software, resolving vulnerabilities in production environments quickly, and capturing the deployment bill of materials (DBOM).
Speakers:
Bob Boule
Robert Boule is a technology enthusiast with PASSION for technology and making things work along with a knack for helping others understand how things work. He comes with around 20 years of solution engineering experience in application security, software continuous delivery, and SaaS platforms. He is known for his dynamic presentations in CI/CD and application security integrated in software delivery lifecycle.
Gopinath Rebala
Gopinath Rebala is the CTO of OpsMx, where he has overall responsibility for the machine learning and data processing architectures for Secure Software Delivery. Gopi also has a strong connection with our customers, leading design and architecture for strategic implementations. Gopi is a frequent speaker and well-known leader in continuous delivery and integrating security into software delivery.
Removing Uninteresting Bytes in Software FuzzingAftab Hussain
Imagine a world where software fuzzing, the process of mutating bytes in test seeds to uncover hidden and erroneous program behaviors, becomes faster and more effective. A lot depends on the initial seeds, which can significantly dictate the trajectory of a fuzzing campaign, particularly in terms of how long it takes to uncover interesting behaviour in your code. We introduce DIAR, a technique designed to speedup fuzzing campaigns by pinpointing and eliminating those uninteresting bytes in the seeds. Picture this: instead of wasting valuable resources on meaningless mutations in large, bloated seeds, DIAR removes the unnecessary bytes, streamlining the entire process.
In this work, we equipped AFL, a popular fuzzer, with DIAR and examined two critical Linux libraries -- Libxml's xmllint, a tool for parsing xml documents, and Binutil's readelf, an essential debugging and security analysis command-line tool used to display detailed information about ELF (Executable and Linkable Format). Our preliminary results show that AFL+DIAR does not only discover new paths more quickly but also achieves higher coverage overall. This work thus showcases how starting with lean and optimized seeds can lead to faster, more comprehensive fuzzing campaigns -- and DIAR helps you find such seeds.
- These are slides of the talk given at IEEE International Conference on Software Testing Verification and Validation Workshop, ICSTW 2022.
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
Welcome to the first live UiPath Community Day Dubai! Join us for this unique occasion to meet our local and global UiPath Community and leaders. You will get a full view of the MEA region's automation landscape and the AI Powered automation technology capabilities of UiPath. Also, hosted by our local partners Marc Ellis, you will enjoy a half-day packed with industry insights and automation peers networking.
📕 Curious on our agenda? Wait no more!
10:00 Welcome note - UiPath Community in Dubai
Lovely Sinha, UiPath Community Chapter Leader, UiPath MVPx3, Hyper-automation Consultant, First Abu Dhabi Bank
10:20 A UiPath cross-region MEA overview
Ashraf El Zarka, VP and Managing Director MEA, UiPath
10:35: Customer Success Journey
Deepthi Deepak, Head of Intelligent Automation CoE, First Abu Dhabi Bank
11:15 The UiPath approach to GenAI with our three principles: improve accuracy, supercharge productivity, and automate more
Boris Krumrey, Global VP, Automation Innovation, UiPath
12:15 To discover how Marc Ellis leverages tech-driven solutions in recruitment and managed services.
Brendan Lingam, Director of Sales and Business Development, Marc Ellis
DevOps and Testing slides at DASA ConnectKari Kakkonen
My and Rik Marselis slides at 30.5.2024 DASA Connect conference. We discuss about what is testing, then what is agile testing and finally what is Testing in DevOps. Finally we had lovely workshop with the participants trying to find out different ways to think about quality and testing in different parts of the DevOps infinity loop.
Smart TV Buyer Insights Survey 2024 by 91mobiles.pdf91mobiles
91mobiles recently conducted a Smart TV Buyer Insights Survey in which we asked over 3,000 respondents about the TV they own, aspects they look at on a new TV, and their TV buying preferences.
Observability Concepts EVERY Developer Should Know -- DeveloperWeek Europe.pdfPaige Cruz
Monitoring and observability aren’t traditionally found in software curriculums and many of us cobble this knowledge together from whatever vendor or ecosystem we were first introduced to and whatever is a part of your current company’s observability stack.
While the dev and ops silo continues to crumble….many organizations still relegate monitoring & observability as the purview of ops, infra and SRE teams. This is a mistake - achieving a highly observable system requires collaboration up and down the stack.
I, a former op, would like to extend an invitation to all application developers to join the observability party will share these foundational concepts to build on:
Encryption in Microsoft 365 - ExpertsLive Netherlands 2024Albert Hoitingh
In this session I delve into the encryption technology used in Microsoft 365 and Microsoft Purview. Including the concepts of Customer Key and Double Key Encryption.
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
Monitoring Java Application Security with JDK Tools and JFR Events
Novel Way to Isolate Adipose Derive Mesenchymal Stem Cells & Its Future Clinical Applications in Obesity
1. IOSR Journal of Biotechnology and Biochemistry (IOSR-JBB)
e-ISSN: XXXX-XXXX, p-ISSN: XXXX-XXXX, Volume 1, Issue 1 (Nov. – Dec. 2014), PP 03-10
www.iosrjournals.org
www.iosrjournals.org 3 | Page
Novel Way to Isolate Adipose Derive Mesenchymal Stem Cells &
Its Future Clinical Applications in Obesity
Eliza Chakraborty1*
And Neelam Shaik2
1,2:
Dept. Of Stem Cell Research and Regenerative Medicine, Vidyapratishthans School of Biotechnology (VSBT,
University of Pune), Baramati Dist-Pune , Pin -413133India.o
Abstract: Adipose-derived stem cells (ADSCs), were isolated from discarded human fat tissue, obtained from c-
section with our recently modified methods, in Stem Cell & Regenerative Medicine Lab, VSBT. Here we develop
two methods to isolate Adipose derived mesenchymal stem cells with enzyme digestion and use of
phosphatidylcholine and deoxycholate. Surface protein expression was analyzed by flow cytometry to
characterize the cell phenotype. The multi-lineage potential of ADSCs was testified by differentiating cells with
adipogenic inducer. ADSCs can be cultured in vitro for up to one month without passage. Also, the flow
cytometry analysis showed that ADSCs expressed high levels of stem cell related surface marker CD105.
ADSCs have strong proliferation ability, maintain their phenotypes, and have stronger multi-differentiation
potential. The molecular basis of ADSC differentiation was studied using bioinformatics tools with an aim to
identify the key proteins involved in differentiation, such that they could be used as potential targets for drug
development for the treatment of obesity. The key proteins involved were found to be PPARG and C/EBPα. The
structures of the proteins were retrieved from MMDB (Molecular Modelling Database) and PDB (Protein Data
Bank) respectively.
Key Words: Adipose-derived stem cells, Mesenchymal stem cells, Enzyme digestion, Phosphatidylcholine,
Deoxycholate, PPARG, C/EBPα, etc.
I. Introduction:
A major source of stem cells is provided by adult bone marrow-derived mesenchymal stem cells
(MSCs), present within the bone marrow stroma. The retrieval of stem cells from the bone marrow, however, is
highly invasive and provides only low numbers of stem cells. (Casteilla et al., 2004, Michelle Locke et al.,
2008) Therefore, alternative sources for MSCs have been investigated. Only recently, adipose tissue was defined
as a new source of MSCs. Like the bone marrow, adipose tissue contains a stromal fraction, which contains a
population of adipose tissue-derived stem cells (ASCs) (Zuk et al., 2001). An advantage of using adipose tissue
as the source of stem cells is that the harvesting of adipose tissue is minimally invasive and can be done under
local anesthesia, thus resulting in minimal patient discomfort and low patient risk. (Michelle Locke et al., 2008)
Furthermore, adipose tissue provides large numbers of stem cells as compared to bone marrow. Adipose-derived
MSC are capable of proliferation in monolayer culture and multi lineage differentiation in response to inductive
conditions, and thus have potential clinical application (Bailey et al., 2010; Fraser et al., 2008; Rigotti et al.,
2009).
The conventional method of isolating MSC from adipose tissue is using lipoaspirate that consists of at
least four main steps: digestion, washing, centrifugation and red blood cell lysis. Adipose tissue from
lipoaspirate samples is incubated with collagenase for up to 1 hour. Then, the digests are washed, and
centrifuged to separate the floating adipocytes from the pelleted stromal cells. The pelleted stromal cells are
finally incubated with red blood cell lysis solution and centrifuged one more time. This procedure generates
tissue fragments that should be removed before cell plating through a 100–150 μm nylon mesh. (Zuk et al.,
2001) Irrespective of the source of tissue, this method is time consuming and expensive, especially when
applied to large volumes of tissue (Baptista et al., 2009); decreased cell viability due to lytic activity is also a
problem with this method (Ishige et al., 2009).
We have described a novel method of isolating MSC from whole adipose tissue. The conventional
methods are focused only on the use of lipoaspirate, but lipoaspirate is sometimes not available for stem cell
isolation and it is also required for reconstructive lipotransfer (R. A. Soriano et al., 2013). Furthermore whole
adipose tissue, in some surgical procedure, like caesarian section etc., has to be removed and discarded as
medical waste. Such adipose tissue obtained after a surgery can be utilized as a source of autologous stem cells.
Our protocol hence is focused on utilization of discarded adipose tissue; it is based on enzymatic tissue digestion
and use of phosphoidylcholine and deoxycholate as fat solubilising agents, so as to prepar lipoaspirate in-vitro.
Despite the major differences between both methods, similar populations of MSC have been isolated. The
populations of cells derived from process were positive for mesenchymal surface marker CD105. They also
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were able to accumulate lipid droplets under specific stimuli for each differentiation event. Their proprieties
support their use for diverse therapeutic applications. Techniques used on these assays will be detailed below.
According to the WHO Factsheet (2014), worldwide obesity has nearly doubled since 1980. In 2008
more than 1.4 billion adults were found to be overweight. Of these over 200 million were male and nearly 300
million females. 65% of world population lives in countries where overweight and obesity kills more people
than underweight. More than 40 million children under age of 5 were overweight or obese in 2012. Childhood
obesity is associated with a high chance of obesity, premature death and disability in adulthood and also
increased future risk of breathing disabilities, hypertension early markers of cardiovascular disease, insulin
resistance and psychological effects.
Overweight and obesity are leading risks for global deaths. Around 3.4 million adults die each year as a
result of being overweight and obese. In addition, 44% of diabetes burden, 23% of ischemic heart disease
burden and between 7% and 41% of certain cancer burdens are attributable to obesity.
Considering these facts we tried to analyze the key proteins involved in differentiation of stem cells to
adipocytes using bioinformatics approach, so as to find a new target for treating obesity. LifeMap DiscoveryTM
database was used along with KEGG pathway database, MMDB and PDB databases to study the differentiation
pathways and protein structure.
The detailed study of databases revealed that two pathways were mainly responsible in stem cell
differentiation viz. insulin pathway and Wnt pathway. It was observed that insulin pathway enhances the
differentiation and wnt pathway inhibits the differentiation. On further analysis the key proteins involved in
differentiation were identified to be PPARG and C/EBPα. C/EBPα is a pleiotropic transcriptional activator of
adipocyte-specific genes. Promoters from numerous adipocyte genes contain C/EBP regulatory consensus
sequences and are trans-activated by C/EBPα. Forced expression of this isoform in 3T3-L1 preadipocytes
stimulates adipogenesis in the absence of any hormonal induction. In addition, blocking its expression wit
antisense C/EBPα RNA inhibits adipogenesis. Furthermore, C/EBPα(−/−) mice have defects in lipid
accumulation. These observations establish the requirement for C/EBPα in preadipocyte differentiation.
(Tamara C. Otto et. al., 2014).
PPARG is a central regulator during adipogenesis. The transcription factor is induced during
differentiation and is responsible for activating a number of genes involved in fatty acid binding, uptake and
storage, including 422/aP2, lipoprotein lipase, acyl coenzyme A synthase and phosphoenol pyruvate
carboxykinase. The importance of PPARG in adipogenesis has been demonstrated in several ways. Over-
expression of PPARG in non adipogenic fibroblasts stimulates adipogenesis. PPARG (−/−) mice die in utero
secondary to a placental defect, but those that survive to term through tetraploid rescue are deficient in brown
fat. PPARG is transcriptionally activated by C/EBPβ and C/EBPδ, and once expressed; PPARG and C/EBPα
positively regulate each other‟s expression. (Tamara C. Otto et.al. 2014)
LifeMap DiscoveryTM
provides investigators with an integrated database of embryonic development,
stem cell biology and regenerative medicine. The hand-curated reconstruction of cell ontology with stem cell
biology; including molecular, cellular, anatomical and disease-related information, provides efficient and easy-
to-use, searchable research tools. The database collates in vivo and in vitro gene expression and guides
translation from in vitro data to the clinical utility, and thus can be utilized as a powerful tool for research and
discovery in stem cell biology, developmental biology, disease mechanisms and therapeutic discovery. LifeMap
Discovery is freely available to academic nonprofit institutions at http://discovery.lifemapsc.com
II. Human adipose tissue mesenchymal stem cells: Chakraborty E. et al’s protocol
Human abdominal subcutaneous adipose tissue was surgically obtained from females after Cesarean
delivery using sterile techniques, with Prior consent of the donors. The sample was first washed in normal saline
and then collected in (1X) PBS containing antibiotic antimycotic (Penicillin and Streptomycin) solution
1ml/100ml and then processed within 5 hours after collection. After transferring under sterile conditions, the
tissue was washed twice in (1X) PBS containing antibiotic antimycotic solution; to remove blood. The tissue
was then cut into two pieces: one of which underwent enzymatic treatment and another was processed for in
vitro lipoaspirate production.
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1. Enzymatic Digestion Method:
Adipose tissue was minced in (1X) PBS using sterile surgical blades and forceps. It was then digested
with same volume of PBS containing 1% BSA and 0.2% Collagenase type IA in a water bath for 60 mins or
more until the solution appeared clear. The solution was then filtered using cell strainer (100µm) to remove
undigested tissue. The filtrate was centrifuged at 1800 RPM for 5 mins. The pellet was resuspended in the same
volume of RBC lysis buffer (160 mM/L NH4Cl in PBS) and incubated for 20 mins at room temperature. The
solution was again centrifuged at 1800 RPM for 5 mins. After centrifugation, the supernatant was discarded and
the ADSC pellet was resuspended in 1ml complete media (DMEM containing 4.5 g/ml glucose, L-glutamine,
sodium pyruvate) with 10% FBS, 1% antibiotic antimycotic solution. Cells were seeded in T25 culture flasks.
3ml of complete medium was added to the flasks. The flasks were incubated in CO2 incubator at 5% CO2, at 37 ͦ
C; for 24 to 48 hours. After 48 hours, 1ml of complete media was added to each flask.
2. In vitro preparation of human lipoaspirate for ADSCs Isolation:
The adipose tissue was minced into small pieces in (1X) PBS using sterile surgical blades and forceps.
The tissue was then incubated with 25mg/ml Phosphatidylcholine(Sigma) and 21mg/ml Deoxycholate(Sigma).
In vitro lipoaspirate was prepared by mixing it by frequent pipetting and then incubating it for 40-60 mins. It
was then digested with same volume of (1X) PBS containing 1% BSA and 0.1% collagenase type IA in a water
bath for 60 mins or more until the solution appeared clear. The solution was then filtered using cell strainer
(100µm) to remove undigested tissue. The filtrate was centrifuged at 1800 RPM for 5 mins. The pellet was
resuspended in the same volume of RBC lysis buffer (160 mM/l NH4Cl in PBS) and incubated for 20 mins at
room temperature. The solution was again centrifuged at 1800 RPM for 5 mins. After centrifugation, the
supernatant was discarded and the ADSC pellet was resuspended in 4ml complete media (DMEM containing
4.5 g/ml glucose, L-glutamine, sodium pyruvate) with 10% FBS, 1% antibiotic antimycotic solution. Cells were
seeded in T25 culture flasks. 3ml of complete medium was added to the flasks. The flasks were incubated in
CO2 incubator at 5% CO2, at 37 ͦ C; for 24 to 48 hours. After 48 hours, 1ml of complete media was added to
each flask.
The change of medium was carried out at the third day, but only half of the medium was replaced.
After 3–5 days full medium change was performed. Next medium changes took place twice a week. The
passaging was performed using 0.25% trypsin-EDTA solution after confluent growth was seen.
3. Adipogenic Differentiation of ADSCs:
Prior to induction, cultured ADSCs were harvested and plated into the desired tissue culture dish, to
attain approximately 80% confluency. The cells were cultured in 2.5ml AdipoXL basal medium (HiMedia)
specific for adipogenic differentiation. Cells were cultured for a minimum of 14 days with appropriate changes
of the medium every 3-4 days. Highly adipogenic ADSCs populations may show signs of lipid vacuole
formation as early as 7 days induction. ADSCs undergoing adipogenic differentiation will develop multiple lipid
vacuoles that may be easily visualized under the light microscope. In addition, adipogenic differentiation was
confirmed using an Oil Red „O‟ staining. Cells were observed under (10X) inverted microscope.
Immunophenotyping of ADSCs by Flow Cytometry (FACS Calibur, BD BioScience), using PE tagged CD105
antibody.
Bioinformatics based study for regulation of adipogenesis was carried out using LifeMap DiscoveryTM
database
(http://discovery.LifeMapsc.com/).
Open the Embryonic Development page.
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www.iosrjournals.org 6 | Page
i. From the drop down select the Organs/Tissues.
ii. Once you are on the page in the search box type Adipose.
iii. Once you track the adipose tissue, click on the tissue card to open the details.
iv. From the options given on the page select the hyperlink to Signals.
v. Developmental signals related to the organ/tissue are listed with their relevant outcomes, developmental
paths and associated cells are given in a tabular format on the page. Signals are listed according to their
order of development from the first ancestor(s).
vi. Select the appropriate signalling pathways responsible for differentiation and study the pathway
accordingly using the links given along.
vii. Analyse and record the key genes responsible for differentiation of the MSCs to adipocytes.
viii. Retrieve the structure of proteins encoded by those genes using the available databases like NCBI or PDB
or MMDB.
ix. Once the structure is retrieved try to find out the ligands that can be used as potential anti obesity drugs.
x. Observations and conclusions were recorded.
III. Result and discussion:
Enzyme digestion and in-vitro lipoaspirate production both proved to be efficient methods for isolation
of stem cells.
Cells were isolated using two different methods: enzymatic digestion and by PC & DC treatment. In
enzyme digestion 0.2% Collagenase type IA was used. Collagenase is unique among proteases in its ability to
degrade the triplehelical native collagen fibrils commonly found in connective tissue. Enzyme digestion
produced a clear solution, from which ADSCs were isolated after further processing. PC & DC treatment of
adipose tissue led to formation of three different layers similar to the layers formed in standard lipoaspirate
production protocol (Patricia A. Zuk et al., 2002)
1. Morphology of adipose-derived cells:
A majority of cells exhibited multiple extensions from a relatively round cell body, whereas a small
number of cells showed a flat and extended cell body with shorter and less obvious extensions.
2. Microscopic observations of stained adipocytes:
ADSCs were differentiated to adipocytes using AdipoXL (HiMedia) basal media. The differentiation of
cells was confirmed by oil red staining.
The differentiation was seen after 8-9 days of induction. The differentiated cells appeared bigger and
round in shape and showed accumulation of multiple lipid vacuoles that appeared red on oil red staining may be
easily visualized under the light microscope.
The differentiation of stem cells into adipocytes confirmed the presence of stem cells in the culture.
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3. Characterisation of ADSCs by FACS analysis:
Fig.8: Dot-plot of labelled sample (PE tagged CD105) and control sample resp.
Fig.9: Histogram obtained from the FACS analysis of PE tagged CD105 labelled sample and control sample
ADSCs were characterized by Flow Cytometry using PE tagged Mouse Anti-human CD105 antibody.
The major shift in the plot in comparison to control, after gating, confirmed the presence of stem cells. CD105 is
a specific surface marker expressed by Adipose derived mesenchymal stem cells. Hence, the presence of CD105
expressing cells confirmed the presence of stem cells.
4. Study of crucial pathways responsible for stem cell differentiation into adipocytes:
The crucial pathways responsible in differentiation were studied using the LifeMap Discovery database.
After exploring the database and studying the given signalling pathways, it was seen that two pathways
play the major role in differentiation of MSCs into adipocytes. The first pathway was, Insulin pathway which
induces the differentiation of MSCs to adipocytes whereas, Wnt pathway inhibits the adipogenesis.
Thus, by inhibiting the insulin pathway and inducing the Wnt pathway the differentiation of adipocytes
can be directed to formation of muscle cells, than the adipocytes; thus, reducing the obesity and enhancing
muscle formation.
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The key proteins regulating the differentiation were found to be the PPARG (peroxisome proliferator-
activated receptor gamma) and C/EBPα. (CCAAT/enhancer binding protein alpha)
C/EBPα is a pleiotropic transcriptional activator of adipocyte-specific genes. Promoters from numerous
adipocyte genes contain C/EBP regulatory consensus sequences and are trans-activated by C/EBPα.
Forced expression of this isoform in 3T3-L1 preadipocytes stimulates adipogenesis in the absence of
any hormonal induction. In addition, blocking its expression wit antisense C/EBPα RNA inhibits adipogenesis.
Furthermore, C/EBPα(−/−) mice have defects in lipid accumulation. These observations establish the
requirement for C/EBPα in preadipocyte differentiation. (Tamara C. Otto and M. Daniel Lane, 2014)
PPARγ is a central regulator during adipogenesis. The transcription factor is induced during
differentiation and is responsible for activating a number of genes involved in fatty acid binding, uptake and
storage, including 422/aP2, lipoprotein lipase, acyl coenzyme A synthase and phosphoenol pyruvate
carboxykinase. The importance of PPARγ in adipogenesis has been demonstrated in several ways. Over-
expression of PPARγ in non adipogenic fibroblasts stimulates adipogenesis. PPARγ (−/−) mice die in utero
secondary to a placental defect, but those that survive to term through tetraploid rescue are deficient in brown
fat. PPARγ is transcriptionally activated by C/EBPβ and C/EBPδ, and once expressed, PPARγ and C/EBPα
positively regulate each other‟s expression. (Tamara C. Otto and M. Daniel Lane, 2014)
7. Nobel Way to Isolate Adipose Derive Mesenchymal Stem Cells & Its Future Clinical Applications ....
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Proteins encoded by these two genes hence can be possible target to control the differentiation of MSCs to
adipocytes.
The structure of the key proteins PPARG and C/EBPα was retrieved from MMDB and PDB respectively.
The anti-bodies or ligands specific to these proteins or RNAi technology can be used as potential treatment for
obesity.
As Obesity is a major health problem resulting from increases in adipocyte size and number, blocking the
differentiation of the MSCs into adipocytes can serve to be an applicable treatment for obesity and also to avoid
obesity in future.
IV. Conclusion:
Lipoaspirate is sometimes not available or cannot be obtained for isolation of stem cells. In some
surgical cases like caesarian section, abdominal surgeries, etc. whole fat tissue is extracted. This tissue is
discarded as medical waste and in such cases this discarded whole tissue can be used for stem cell isolation. In
contrast to the conventional method that makes use of lipoaspirate, the use of whole adipose tissue can prove to
be useful. With our present method we can preserve and banking adult stem cell also from adult person‟s future
use which can help them to treat Diabetes and other diseases by using a very tiny amount of adipose tissue.
The key proteins involved in stem cell differentiation were identified using bioinformatics tools and
biological databases, LifeMap DiscoveryTM
database and KEGG pathway database. The key proteins playing a
crucial role in differentiation were identified to be PPARG and C/EBPα.
Thus, by blocking these proteins, it would be possible to control the adipogenesis of stem cells. The
anti-bodies or ligands raised against these proteins can be used as potential drugs for treating obesity and may
also help in avoiding obesity in future. Also, RNAi technology can be used for blocking the translation of the
genes coding these proteins.
Acknowledgement:
I would like to express my deep and sincere gratitude to our Dr. Sushama Chaphalkar, Director, VSBT for
giving us an opportunity to work on this project and her dynamism, vision, sincerity and motivation have deeply
inspired us.
I am very grateful to VSBT, for funding the project.
I sincerely thank Dr. Amit Gupta, for his valuable help in operation of FACS machine.
References:
[1]. Ali M. Sharifi, Roshanak Ghazanfari, Neda Tekiyehmaroof, Mohammad A. Sharifi. (2014) Isolation, Cultivation, Characterization
and Expansion of Human Adipose-derived Mesenchymal Stem Cell for Use in regenerative Medicine. International Journal of
Hematology Oncology and Stem Cell Research, 6:1, 1-5.
[2]. Bruno Machado Bertassoli1, Antonio Chaves de Assis Neto, Franceliusa Delys de Oliveira, Maria Angelica Machado Arroyo,
Juliana Shimara Pires Ferrao, Jodonai Barbosa da Silva, Graciela Conceicao Pignatari, and Patrícia Beltrao Braga. (2013)
Mesenchymal Stem Cells - Emphasis in Adipose Tissue. Braz. Arch. Biol. Technol, 56:4, 607-617.
Fig.12: Image showing the role of Wnt pathway
in adipocytes‟ differentiation.
Fig.13: Image showing the structure of PPARG,
retrieved from MMDB (MMDB ID-66390)
Fig.14: Image showing the structure of
C/EBPα , retrieved from PDB.
Fig.11: Image showing the role of Insulin
pathway in adipocytes differentiation and the
key genes involved in it.
8. Nobel Way to Isolate Adipose Derive Mesenchymal Stem Cells & Its Future Clinical Applications ....
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[3]. Catherine M. Kolf, Elizabeth Cho, and Rocky S. Tuan. (2007) Biology of adult mesenchymal stem cells: Regulation of niche, self-
renewal and differentiation. Arthritis Research & Therapy, 9:204.
[4]. Dominic N. Reeds, B. Selma Mohammed, Samuel Klein, Craig Brian Boswell, and V. Leroy Young. Metabolic and Structural
Effects of Phosphatidylcholine and Deoxycholate Injections on Subcutaneous Fat: A Randomized, Controlled Trial. Aesthetic
Surgery Journal, 33:3, 400-408.
[5]. Doris Maria Hexsel, Marcio Serra, Taciana de Oliveira Dal‟Forno, and Debora Zechmeister do Prado. (2005) Cosmetic Uses of
Injectable Phosphatidylcholine on the Face. Otolaryngol Clin N Am, 38:1119-1129.
[6]. Fabian Mohra, Konstanze Deohnerb, Christian Buskea, and Vijay P.S. Rawata. (2011) TET Genes: new players in DNA
demethylation and important determinants for stemness. Experimental Hematology, 39:272-281.
[7]. Giovanni Salti, Ilaria Ghersetich, Franca Tantussi, Bruno Bovani, and Torello Lotti. (2008) Phosphatidylcholine and Sodium
Deoxycholate in the Treatment of Localized Fat: A Double-Blind, Randomized Study. Dermatology Surgery, 34:60-66.
[8]. Hakan Orbay, Morikuni Tobita, and Hiroshi Mizuno. (2012) Mesenchymal Stem Cells Isolated from Adipose and Other Tissues:
Basic Biological Properties and Clinical Applications. Stem Cells International, 9.
[9]. Kamshoushy A., Abel Maksoud R., and Mahdy N. (2012) Evaluation of the Efficacy of Injection Lipolysis using
Phosphatidylcholine/Deoxycholate Versus Deoxycholate Alone in Treatment of Localized Fat Deposits. Journal of Clinical
Experiments of Dermatology, 3:2.
[10]. Khalid Mahmud and Charles E. Crutchfield. (2012) Lipodissolve for Body Sculpting Safety, Effectiveness, and Patient Satisfaction.
Journal of Clinical Aesthetic Dermatology, 5:16-19.
[11]. Kyung Rok Yu and Kyung Sun Kang. (2013) Aging-Related Genes in Mesenchymal Stem Cells: A Mini-Review. Gerontology,
59:557-563.
[12]. Luisa Cimmino, Omar Abdel Wahab, Ross L. Levine, and Iannis Aifantis. (2011) TET Family Proteins and Their Role in Stem Cell
Differentiation and Transformation. Stem Cell, 9.
[13]. Maikel J. Oedayrajsingh Varma, Roel G. M. Breuls, Tabitha E. Schouten, Wouter J.F.M. Jurgens, Hetty J. Bontkes, Gerrit J.
Schuurhuis, S. Marieke Vam Ham, and Florine J. Van Milligen. (2007) Phenotypical and Functional Characterization of Freshly
Isolated Adipose Tissue-Derived Stem Cells. Stem Cells and Development, 16:91-104.
[14]. Mark Palmer, John Curran, and Patrick Bowler. Clinical experience and safety using phosphatidylcholine injections for the
localized reduction of subcutaneous fat: a multicentre, retrospective UK study. Journal of Cosmetic Dermatology, 5:218-226.
[15]. Michelle Locke, John Windsor, and P. Rod Dunbar. (2009) Human adipose-derived stem cells: isolation, characterization and
applications in surgery. ANZ J Surg, 79:235-244.
[16]. Michael P. Francis, Patrick C. Sachs, Lynne W. Elmore, and Shawn E. Holt. (2010) Isolating adipose-derived mesenchymal stem
cells from lipoaspirate blood and saline fraction. Organogenesis, 6:1, 11-14.
[17]. Miguel Ramalho Santos, Soonsang Yoon, Yumi Matsuzaki, Richard C. Mulligan, and Douglas A. Melton. (2002) “Stemness”:
Transcriptional Profiling of Embryonic and Adult Stem Cells. Science, 298.
[18]. Min Zhu, Sepideh Heydarkhan-Hagvall, Marc Hedrick, Prosper Benhaim, and Patricia Zuk. (2013) Manual Isolation of Adipose-
derived Stem Cells from Human Lipoaspirates. Journal of Visual Experiments, 79:1-10.
[19]. N. K. Singh, A.M.M.T. Reza and S. Subi. (2013) Keratinocyte growth factor alters signaling factors to transform mammary fat pad
adipose stem cells isolated from Korean black goat to epithelium. Journal of Veterinary Science & Technology, 4:4.
[20]. Patricia A. Zuk, Min Zhu, Hiroshi Mizuno, Jerry Huang, J. William Futrell, Adam J. Katz, Prosper Benhaim, H. Peter Lorenz, and
Marc H. Hedrick. (2001) Multilineage Cells from Human Adipose Tissue: Implications for Cell-Based Therapies. Tissue
Engineering, 7:2.
[21]. Patricia A. Zuk, Min Zhu, Peter Ashjian, Daniel A. De Ugarte, Jerry I. Huang, Hiroshi Mizuno, Zeni C. Alfonso, John K. Fraser,
Prosper Benhaim, and Marc H. Hedrick. (2002) Human Adipose Tissue Is a Source of Multipotent Stem Cells. Molecular Biology
of the Cell, 13:4279-4295.
[22]. Phanette Gir, Georgette Oni, Spencer A. Brown, Ali Mojallal, and Rod J. Rohrich. (2012) Human Adipose Stem Cells: Current
Clinical Applications. Plast. Reconstr. Surg. 129: 1277.
[23]. Robert R. Bowers and M. Daniel Lane. (2008) Wnt signaling and adipocyte lineage commitment. Cell Cycle 7:9, 1191-1196.
[24]. R. A. Soriano, H. Lamblet, S. A. Mohammadi, and H. Torfi. (2013) Optimization of Roche Liberase TM Research Grade in the
Enzymatic Digestion of Human Adipose Tissue for the Isolation of Stem and Regenerative Cells. Invitrx Therapeutics.
[25]. Scott Karolchyk and Linda Seibert. Detergent Effects of Sodium Deoxycholate Are a Major Feature of an Injectable
Phosphatidylcholine Formulation Used for Localized Fat Dissolution: A Review. American Journal of Mesotherapy.
[26]. Silvana Baglioni, Michela Francalanci, Roberta Squecco, Adriana Lombardi, Giulia Cantini, Roberta Angeli, Stefania Gelmini,
Daniele Guasti, Susanna Benvenuti, Francesco Annunziato, Daniele Bani, Francesco Liotta, Fabio Francini, Giuliano Perigli, Mario
Serio, and Michaela Luconi. (2009) Characterization of human adult stem-cell populations isolated from visceral and subcutaneous
adipose tissue. The FASEB Journal.
[27]. Stephen R. Farmer. (2006) Transcriptional control of adipocyte formation. Cell Metab, 4:4, 263-273.
[28]. Tamara C. Otto and M. Daniel Lane. (2014) Adipose Development: From Stem Cell to Adipocyte. Critical Reviews in
Biochemistry and Molecular Biology.
[29]. Yangsik Jeong and David J. Mangelsdorf. (2009) Nuclear receptor regulation of stemness and stem cell Differentiation.
Experimental and Molecular Medicine, 41:8, 525-537.
[30]. Xu Fang Yang, Xu He, Jian He, Li Hong Zhang, Xue Jin Su, Zhi Yong Dong, Yun Jian Xu, Yan Li, and Yu Lin Li. (2011) High
efficient isolation and systematic identification of human adipose-derived mesenchymal stem cells. Journal of Biomedical Science,
18:59.
[31]. Yu Ren, Haiqing Wu, Xueyuan Zhou, Jianxun Wen, Muzi Jin, Ming Cang, Xudong Guo. (2012) Isolation, expansion, and
differentiation of goat adipose-derived stem cells. Research in Veterinary Science, 93:404-411.