This document summarizes a presentation about a novel "knock-in" mouse model that expresses a catalytically inactive form of tissue transglutaminase (TG2). The TG2-C277S mouse model was developed using CRISPR/Cas9 gene editing to mutate the active site cysteine residue of TG2. Biochemical analysis showed the mutant TG2 lacked crosslinking activity but retained GTP binding ability. Preliminary analysis found the TG2-C277S mice expressed normal levels of TG2 protein and its interaction with integrins and fibronectin were preserved. This novel mouse model will provide insights into TG2 functions in vascular aging independent of its enzymatic crosslinking activity.
Learn about the power of Regenerative Medicine or Orthobiologics. Engage the science on how stem cells and platelet rich plasma can improve quality of life and function in orthopedic needs. Check out more at www.JaxStemcell.com
Forces associated with blood flow are major determinants of vascular morphogenesis and physiology. Blood flow is crucial for blood vessel development during embryogenesis and for regulation of vessel diameter in adult life. It is also a key factor in atherosclerosis, which, despite the systemic nature of major risk factors, occurs mainly at regions of arteries that experience disturbances in fluid flow. Recent data have highlighted the potential endothelial mechanotransducers that mediate responses to flow, the effects of atheroprotective versus atherogenic flow, and the mechanisms that contribute to progression of the disease over time and how systemic factors interact with flow patterns to cause atherosclerosis.
Frm:- Nat Rev Mol Cell Biol. 2009 Jan; 10(1): 53–62. doi: 10.1038/nrm2596
Cardiac Inflammation and Repair Following Myocardial InfarctionInsideScientific
Join Dr. Merry Lindsey as she discusses her research involving the physiology of recovery from cardiac events.
Age plays a pivotal role in the deterioration of cardiovascular functionality, resulting in an increased risk of cardiovascular disease in older adults. The prevalence of cardiovascular disease has also been shown to increase with age, in both men and women, including the prevalence of atherosclerosis, stroke and, myocardial infarction.
Following myocardial infarction (MI), the left ventricle (LV) undergoes a series of cardiac wound healing responses that involve both the stimulation of robust inflammation to clear necrotic myocytes and tissue debris and the induction of extracellular matrix (ECM) protein synthesis to generate an infarct scar. Collectively, this process in known as LV remodeling. Matrix metalloproteinase-9 (MMP-9) is a key regulator of LV remodeling post-MI, through direct effects on ECM turnover as well as indirect effects on the regulation of the major cell types that coordinate cardiac wound healing- namely the infiltrating leukocytes and the cardiac fibroblasts. We will discuss recent research that has expanded our understanding of MI LV remodeling, including recent proteomic advances focused on the ECM compartment to provide novel functional and translational insights. In summary, this webinar will provide an overview of how cardiac ECM research has evolved over the last decade and will provide insight into future directions that will drive further understanding of MMP directed cardiac ECM turnover after MI.
Adult Stem cells in Orthopaedics present and future perspectives.
Παρουσίαση του Δρ. Σταύρου Αλευρογιάννη που έγινε στο ξενοδοχείο Χίλτον, στις 12/06/15 στα πλαίσια Ημερίδας της Ελληνικής Εταιρείας Αναγεννητικής Ιατρικής, Αντιγήρανσης και Βιοτεχνολογίας, στο 41ο Πανελλήνιο Ιατρικό Συνέδριο.
"H θέση της αναγεννητική Ιατρικής στις παθήσεις Οστών και Αρθρώσεων"
Learn about the power of Regenerative Medicine or Orthobiologics. Engage the science on how stem cells and platelet rich plasma can improve quality of life and function in orthopedic needs. Check out more at www.JaxStemcell.com
Forces associated with blood flow are major determinants of vascular morphogenesis and physiology. Blood flow is crucial for blood vessel development during embryogenesis and for regulation of vessel diameter in adult life. It is also a key factor in atherosclerosis, which, despite the systemic nature of major risk factors, occurs mainly at regions of arteries that experience disturbances in fluid flow. Recent data have highlighted the potential endothelial mechanotransducers that mediate responses to flow, the effects of atheroprotective versus atherogenic flow, and the mechanisms that contribute to progression of the disease over time and how systemic factors interact with flow patterns to cause atherosclerosis.
Frm:- Nat Rev Mol Cell Biol. 2009 Jan; 10(1): 53–62. doi: 10.1038/nrm2596
Cardiac Inflammation and Repair Following Myocardial InfarctionInsideScientific
Join Dr. Merry Lindsey as she discusses her research involving the physiology of recovery from cardiac events.
Age plays a pivotal role in the deterioration of cardiovascular functionality, resulting in an increased risk of cardiovascular disease in older adults. The prevalence of cardiovascular disease has also been shown to increase with age, in both men and women, including the prevalence of atherosclerosis, stroke and, myocardial infarction.
Following myocardial infarction (MI), the left ventricle (LV) undergoes a series of cardiac wound healing responses that involve both the stimulation of robust inflammation to clear necrotic myocytes and tissue debris and the induction of extracellular matrix (ECM) protein synthesis to generate an infarct scar. Collectively, this process in known as LV remodeling. Matrix metalloproteinase-9 (MMP-9) is a key regulator of LV remodeling post-MI, through direct effects on ECM turnover as well as indirect effects on the regulation of the major cell types that coordinate cardiac wound healing- namely the infiltrating leukocytes and the cardiac fibroblasts. We will discuss recent research that has expanded our understanding of MI LV remodeling, including recent proteomic advances focused on the ECM compartment to provide novel functional and translational insights. In summary, this webinar will provide an overview of how cardiac ECM research has evolved over the last decade and will provide insight into future directions that will drive further understanding of MMP directed cardiac ECM turnover after MI.
Adult Stem cells in Orthopaedics present and future perspectives.
Παρουσίαση του Δρ. Σταύρου Αλευρογιάννη που έγινε στο ξενοδοχείο Χίλτον, στις 12/06/15 στα πλαίσια Ημερίδας της Ελληνικής Εταιρείας Αναγεννητικής Ιατρικής, Αντιγήρανσης και Βιοτεχνολογίας, στο 41ο Πανελλήνιο Ιατρικό Συνέδριο.
"H θέση της αναγεννητική Ιατρικής στις παθήσεις Οστών και Αρθρώσεων"
Monocytes and macrophages are innate immune cells that reside and accumulate in atherosclerotic lesions but also in the healthy and injured heart and brain. The cells and their subsets pursue distinct functions in steady state and disease, and their tenure may range between hours to months. Some subsets are highly inflammatory, while others support tissue repair.
Dr. Matthias Nahrendorf discusses current concepts of cell supply by the hematopoietic system, lineage relationships and systems’ cross talk, highlights open questions, and describes imaging tools for studying monocytes, macrophages and their progenitors.
Key Topics Include:
- Resident versus bone marrow derived macrophages
- Roles and phenotypes of heart leukocytes
- Hematopoiesis and the bone marrow in cardiovascular disease
Regenerative medicine is now an recognized specialty which has evolved from degerative diseases of Orthopaedic Surgery.Orthobiologics is a current terminology for the application of various cells, cytokines, growth factors.Busy people find it to update and this is an update.
Objective: Ischemia-reperfusion (I/R) leads to reactive oxygen species formation and cell death in kidney tissue with injury and organ transplantation. Simvastatin (SIM) is an antioxidant, anti-inflammatory, and anticoagulant agent. Alterations in I/R-induced acute kidney injury model with SIM treatment were analyzed.
Study Design: Wistar rats (n=28) were grouped into Sham, Ischemia, I/R, and I/R+SIM treated. Left rat kidney renal vessels were clamped for 60 minutes for ischemia, and the I/R group had 6 hours of reperfusion. 10 mg/kg SIM was given orally for 28 days. MDA, GSH, and MPO were analyzed. Kidney tissues were paraffin embedded, and primary antibodies TNF-α and caspase-3 were applied for immunohistochemistry.
Results: In the I/R group, intense inflammatory cell infiltration around the vessels and necrosis in the glomerular structures were observed. In the treated group, proximal and distal tubular cells were found to be close to normal. Immunoexpression of caspase-3 in the ischemia group was positive in degenerative glomeruli. In the treated group, TNF-α expression was negative in the glomerular structures. MDA and MPO levels were significantly increased in ischemia and I/R.
Conclusion: We suggest that SIM treatment improved kidney tissue structure and function in a model of I/R injury.
Keywords: caspase-3; immunohistochemistry; ischemia/reperfusion; kidney; MPO; simvastatin
Monocytes and macrophages are innate immune cells that reside and accumulate in atherosclerotic lesions but also in the healthy and injured heart and brain. The cells and their subsets pursue distinct functions in steady state and disease, and their tenure may range between hours to months. Some subsets are highly inflammatory, while others support tissue repair.
Dr. Matthias Nahrendorf discusses current concepts of cell supply by the hematopoietic system, lineage relationships and systems’ cross talk, highlights open questions, and describes imaging tools for studying monocytes, macrophages and their progenitors.
Key Topics Include:
- Resident versus bone marrow derived macrophages
- Roles and phenotypes of heart leukocytes
- Hematopoiesis and the bone marrow in cardiovascular disease
Regenerative medicine is now an recognized specialty which has evolved from degerative diseases of Orthopaedic Surgery.Orthobiologics is a current terminology for the application of various cells, cytokines, growth factors.Busy people find it to update and this is an update.
Objective: Ischemia-reperfusion (I/R) leads to reactive oxygen species formation and cell death in kidney tissue with injury and organ transplantation. Simvastatin (SIM) is an antioxidant, anti-inflammatory, and anticoagulant agent. Alterations in I/R-induced acute kidney injury model with SIM treatment were analyzed.
Study Design: Wistar rats (n=28) were grouped into Sham, Ischemia, I/R, and I/R+SIM treated. Left rat kidney renal vessels were clamped for 60 minutes for ischemia, and the I/R group had 6 hours of reperfusion. 10 mg/kg SIM was given orally for 28 days. MDA, GSH, and MPO were analyzed. Kidney tissues were paraffin embedded, and primary antibodies TNF-α and caspase-3 were applied for immunohistochemistry.
Results: In the I/R group, intense inflammatory cell infiltration around the vessels and necrosis in the glomerular structures were observed. In the treated group, proximal and distal tubular cells were found to be close to normal. Immunoexpression of caspase-3 in the ischemia group was positive in degenerative glomeruli. In the treated group, TNF-α expression was negative in the glomerular structures. MDA and MPO levels were significantly increased in ischemia and I/R.
Conclusion: We suggest that SIM treatment improved kidney tissue structure and function in a model of I/R injury.
Keywords: caspase-3; immunohistochemistry; ischemia/reperfusion; kidney; MPO; simvastatin
EUTOX CME ERA-EDTA 2016 Vienna
A short overview on unhealthy endothelium during CKD and the role of selected uremic toxins mainly indoles, but with chocolate and zebrafish inside.
ANS Testing Device University of Miami Study PresentationMaxiMedRx
Visit www.maximedrx.com
Sudopath Sudoscan testing device diabetic study by university of miami. Evaluating a New Approach to Detect Type 2 Diabetes Mellitus using the ES Complex-TSS
Learning Objectives:
Describe the consequences of hyper- and hypovolemia for surgical and critically ill patients.
Develop a fluid management strategy for individual patients
Aterosclerosi il danno d'organo: vasculopatia periferica - di P. Buonamico MedOliveOil
Aterosclerosi il danno d'organo: vasculopatia periferica - di P. Buonamico. 7 giugno 2012. Corso di formazione "valore nutrizionale e salutistico di prodotti agroalimentari” - Università degli studi di Bari.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
This presentation highlights the applications and capabilities of the M-Series™ compact MRI systems. Anatomical, functional, and molecular imaging can be performed on the M-Series and are often applied in cancer, cardiac, neuroscience, and multimodal imaging studies. It showcases example data from a variety of papers and training sessions in which the focus is on anatomy, neurobiology, and oncology. The presentation shows data from contrast agents which further enhances the capabilities of the M-Series, providing invaluable insights into tissue/tumor perfusion, myocardial infarction size, and molecular targets.
Ultrasound color Doppler imaging has been routinely used for the diagnosis of cardiovascular diseases, enabling real-time flow visualization through the Doppler effect. Yet, its inability to provide true flow velocity vectors due to its one-dimensional detection limits its efficacy. To overcome this limitation, various VFI schemes, including multi-angle beams, speckle tracking, and transverse oscillation, have been explored, with some already available commercially. However, many of these methods still rely on autocorrelation, which poses inherent issues such as underestimation, aliasing, and the need for large ensemble sizes. Conversely, speckle-tracking-based VFI enables lateral velocity estimation but suffers from significantly lower accuracy compared to axial velocity measurements.
To address these challenges, we have presented a speckle-tracking-based VFI approach utilizing multi-angle ultrafast plane wave imaging. Our approach involves estimating axial velocity components projected onto individual steered plane waves, which are then combined to derive the velocity vector. Additionally, we've introduced a VFI visualization technique with high spatial and temporal resolutions capable of tracking flow particle trajectories.
Simulation and flow phantom experiments demonstrate that the proposed VFI method outperforms both speckle-tracking-based VFI and autocorrelation VFI counterparts by at least a factor of three. Furthermore, in vivo measurements on carotid arteries using the Prodigy ultrasound scanner demonstrate the effectiveness of our approach compared to existing methods, providing a more robust imaging tool for hemodynamic studies.
Learning objectives:
- Understand fundamental limitations of color Doppler imaging.
- Understand principles behind advanced vector flow imaging techniques.
- Familiarize with the ultrasound speckle tracking technique and its implications in flow imaging.
- Explore experiments conducted using multi-angle plane wave ultrafast imaging, specifically utilizing the pulse-sequence mode on a 128-channel ultrasound research platform.
Accelerating the Delivery of New Treatments for Children with Neuroblastoma 2...Scintica Instrumentation
Neuroblastoma is a tumour arising from anomalies in the development of the sympathic nervous system and still accounts for 13% of all cancer-related death in children due to resistant, relapsing and metastatic diseases. There is an urgent need for the development of new treatment against high-risk relapsed neuroblastoma.
Overview:
Here we will discuss the ICR Paediatric Mouse Hospital approach which integrates more advanced mouse modelling, such as the use of genetically-engineered mouse (GEM) models and patient-derived xenografts to accelerate the discovery and evaluation of novel therapeutic strategies and help shape the clinical trial pipeline priorities for children with high-risk relapsing/refractory neuroblastoma.
We will also highlight the pivotal role of MRI within the Mouse Hospital which includes:
Enhancing and accelerating preclinical trials
Quantitatively inform on tumour phenotype and tumour response to treatment to:
Develop in vivo models that emulate the clinical treatment resistant phenotype using chemotherapy-dose escalation protocol
Characterize tumour spatial heterogeneity and evolution over treatment and guide the pathological and molecular characterization of the resistant phenotype
Finally we will also discuss how the compact, cryogen-free and user-friendly Aspect Imaging M-Series has transformed our way of working within the mouse hospital by providing a shared and easily accessible resource for tumour screening (with minimal onboarding) .
(March 14, 2024) Webinar: Validation of DEXA for Longitudinal Quantification ...Scintica Instrumentation
Noninvasive imaging is central to preclinical, in vivo models of pancreatic ductal adenocarcinoma (PDAC). While bioluminescent imaging (BLI) is a gold standard, its signal is dependent on the metabolic activity of tumor cells. In contrast, dual energy X-ray absorptiometry (DEXA) is a direct measure of body composition. Thus, this project aimed to assess the potential of using DEXA for longitudinal quantification of tumor burden versus BLI in an orthotopic KCKO murine model of PDAC. In short, DEXA successfully identified a growing tumor burden and accurately predicts ex vivo tumor mass in a time sensitive manner.
Learning objectives:
Learn to take advantage of DEXA for things other than bone density and bone health (i.e., lean, and fat mass)
Understand that DEXA can reproducibly and accurately be used to monitor tumor burden and growth in orthotopic murine models of pancreatic cancer
Understand the importance of repurposing techniques and equipment for new analysis
Understand that non-invasive in vivo imaging is crucially important in severely compromised models like those for PDAC and other cancers
See the value of utilizing multiple techniques throughout an experiment to enhance data collection
(March 13, 2024) Overview of Preclinical Small Animal and Multimodal ImagingScintica Instrumentation
In this webinar, we reviewed some of the most commonly used preclinical imaging modalities, including magnetic resonance imaging (MRI), positron emission tomography (PET), computer tomography (CT), ultrasound, photoacoustic, bioluminescence, fluorescence, dual-energy x-ray absorptiometry (DEXA/DXA), and intravital microscopy. For each modality, we spent time reviewing the basics of how each worked, the strengths and considerations of each, and some key application areas and example images. Finally, we discussed the benefits of multimodal imaging and reviewed a few papers utilizing a variety of imaging modalities to help support their research outcomes.
We ended with a very brief introduction to Scintica Instrumentation and our philosophy behind the various products we represented. However, the main focus of the webinar was on education, and not our diverse product portfolio.
Dr. Lawrence Yip explained how Photoacoustic (PA) imaging works, where it fits in with other modalities and, how your research could benefit from this emerging technology.
Excellent spatial resolution, depth penetration, and superior contrast are just some of the advantages often associated with PA imaging. In this webinar, we dove into the advantages, where they can be beneficial, and how the TriTom’s patented technology overcomes some of the challenges experienced by early adopters of this imaging modality.
The TriTom is a turnkey, compact, tabletop imaging system that combines the sensitivity of fluorescence molecular tomography with the depth penetration and spatial resolution of PA tomography. Many applications including cancer, neuroimaging, developmental biology, and cardiovascular research could benefit from adding these imaging modalities, and we will draw from literature and concrete examples to demonstrate this advantage.
Overview:
In this webinar, Dr. Edwin C. Pratt discussed the realm of positron emission tomography (PET) imaging and explained the innovative concept of multiplexed PET. This new scientific advancement makes it possible to perform simultaneous imaging with two different isotopes providing more in depth information with a single scan.
Key Takeaways:
Multiplexed PET is a new reconstruction method to identify and separate positron from positron-prompt gamma emissions without new hardware from list mode PET scanners or energy discrimination of events.
Multiplexed PET is a quantitative method that is agnostic to the type of radiotracer used (IE no compartment modeling). Only a simple uniformity and sensitivity phantom is required.
Acquisition has been shown in a variety of preclinical and clinical PET scanners, though not all scanners can natively acquire data for multiplexing.
Multiplexed PET enables faster throughput for screening radiotracers, or conversely two tracer information of a tissue of interest, like imaging the tumor microenvironment for two immune populations.
(June 29, 2023) Webinar: Designer and Targeted Contrast Agent for Photoacoust...Scintica Instrumentation
Overview:
The talk focused on the synthesis, characterization and use of a novel contrast agent composed of indocyanine green dye for NIR-I photoacoustic (PA) imaging. The contrast agent can be easily tuned to different sizes without enclosure in nanocarriers, has strong optical absorption and PA signal at 895 nm, can be easily functionalized with different targeting molecules and can be imaged for 120 minutes in vivo. The presentation explained details on the genesis of the idea for building a biocompatible contrast agent and give details on its easy synthesis protocols, touch upon a functionalization scheme for adding targeting molecules and demonstrate its use as a PA contrast in mice using the TriTom small animal imaging system.
Photoacoustic imaging (PAI) is a noninvasive imaging modality that relies on absorption of laser light and thermal expansion of biological tissues, which generate ultrasonic waves. These ultrasound waves are then used to reconstitute an image of the tissues with anatomical details and functional information. To increase imaging depth and resolution, PAI requires exogenous molecular contrast agents with high optical absorption in the near infrared (NIR). However, the current repository of NIR dyes that are suitable for PAI is extremely limited. The FDA-approved indocyanine green (ICG) is the only commercially available contrast agent with NIR absorbance that is already used for PAI. However, ICG dyes suffer from poor photostability and high clearance rate.
In this webinar, Dr. Shrishti Singh presented a synthesis method for clinically translatable ICG-JA whose mean size can be finely tuned from 200 nm to 1000 nm and that does not require encapsulation in a nanocarrier. The talk will also detail complete characterization of the agent and steps for functionalization with targeting peptides or antibodies. Additionally, the webinar also provided details about the PA properties of the contrast in vitro in different conditions including whole blood, followed by details on the photoacoustic imaging in vivo using the TriTom system.
Learning Objectives:
Get details on the synthesis of a NIR contrast without the need of a nanocarrier.
Learn in detail about what characteristics a contrast agent should possess to qualify as a clinically translatable technology.
Become familiar with methods to create a targeted contrast agent.
Overview:
In this webinar, Dr. David Maridas from Harvard School of Medicine will shared how the latest Dual X-ray Absorptiometry (DXA) advances helped his research. Dr. Maridas will walked us through comparing different systems used in acquiring and analyzing data.
Dual X-ray absorptiometry allows measurements of fat mass, lean mass, bone mineral density, and bone mineral content in live animals. The short radiation exposure time allows for measurements to be performed in vivo using relatively little anesthesia (e.g., isoflurane), making DXA a powerful tool to frequently measure multiple parameters throughout the life of an animal.
Over the years, multiple DXA systems were implemented with different strengths and weaknesses. During this webinar, we will compared the PIXImus and the iNSiGHT for measurements in adult mice.
Looking at the latest advancements in the field of Dual X-ray Absorptiometry (DXA), Dr. Maridas focused on the comparison of two systems, the iNSiGHT and the PIXImus, which are important tools for the accurate measurement of fat mass, lean mass, bone mineral density, and bone mineral content in live animals.
He critically evaluated and contrast the strengths and weaknesses of these systems in terms of their functionalities, logistics, and the unique challenges they pose, providing guidance for researchers in making the most suitable choice based on their specific requirements and constraints. In conclusion, the selection of a system would depend on the need of scientists and the available space for the machine.
Key Points:
-Evolution of DXA Technology: Watch & learn about the technological advancements in DXA as Dr. Maridas delves into the transformation from legacy to contemporary systems, illustrating how the development of this technology impacts the precision and efficiency of live animals.
-iNSiGHT-ful Comparison: With first-hand experience working with both the iNSiGHT and the PIXImus DXA systems, Dr. Maridas will provided an in-depth, comparative analysis, presenting unique strengths and functionalities.
-Behind-the-Scenes Coordination: Gain exclusive insights into the dynamics and understand how the systems influenced Dr. Maridas’s research outcomes.
(May 3, 2023) Webinar: Exploring a Novel NIR-2 Photoacoustic Agent to Improve...Scintica Instrumentation
In this webinar, we explored the development and characterization of a novel photoacoustic (PA) contrast agent suitable for in vivo applications, that out-performs current conventional PA agents. Translation of transabdominal PA imaging is limited due to high optical attenuation of tissue when imaging at conventional wavelengths from the first near infrared (NIR) window. Though imaging at the NIR-2 window allows deeper light penetration due to minimized tissue scattering, there is a lack of optical contrast from endogenous chromophores at these wavelengths.
Currently, gold nanorods (AuNRs) are conventionally used as exogenous contrast for PA imaging at the NIR-2 window. However, AuNRs have large sizes leading to poor clearance in biological systems with adverse long-term effects due to bioaccumulation. Hence, there is a need for exogenous agents with high optical contrast at NIR-2 suited for in vivo applications. Here, using the TriTom compact PA tomography system, a semiconductor nanocrystal contrast agent was validated in vitro, then demonstrated in a mouse model.
Key Points:
-Explore enhancement in photoacoustic image contrast using a novel NIR-2 agent
-Compare increase in image contrast using the novel NIR-2 agent against conventional exogenous photoacoustic contrast agents
(April 5, 2023) Webinar: Prodigy Open-Platform Research Ultrasound System Ov...Scintica Instrumentation
Overview:
In this webinar, we provided an overview of the Prodigy open-platform research ultrasound system. The Prodigy by S-Sharp is a flexible and powerful ultrasound platform enabling research in ultrasound imaging, high-intensity focused ultrasound (HIFU), non-destructive testing (NDT), and much more. Sold for many years as an OEM component of other systems (e.g., for photoacoustic imaging), this highly capable system is now available to laboratories and researchers around the world.
This compact, high-performance ultrasound system is optimized for a variety of engineering research applications. As an open platform research ultrasound system, the Prodigy allows almost every aspect of ultrasound generation and detection to be customized. This includes true arbitrary transmit waveforms, super-fast acquisition capabilities, rapid data transfer, and a software backend that allows for real-time access and processing of both raw and beamformed data.
Some highlights of the Prodigy include its capability for true arbitrary transmit waveforms by using linear amplifiers with digital-to-analog converters (DAC) and the availability of a graphic user interface for designing pulse sequences and adjusting transmit/receive parameters.
Learn the capabilities of this flexible system with peer-reviewed examples of its many possible applications.
Key Points:
(April 4, 2023) Overview of Preclinical Small Animal Imaging Modalities & Mul...Scintica Instrumentation
Overview:
In this webinar, we will review some of the most used preclinical imaging modalities, including magnetic resonance imaging (MRI), positron emission tomography (PET), computer tomography (CT), ultrasound, photoacoustic, bioluminescence, fluorescence, dual-energy x-ray absorptiometry (DEXA/DXA) and intravital microscopy. For each modality, we will spend time reviewing the basics of how each works, the strengths and considerations of each, and some key application areas and example images. Finally, we will discuss the benefits of multimodal imaging and review a few papers utilizing a variety of imaging modalities to help support their outcomes. This webinar will introduce our educational focus on preclinical imaging modalities coming up in 2023.
The webinar will be a brief introduction for those who need to become more familiar with all or some of the preclinical imaging modalities. At the same time, our educational focus over the year will dive deeper into each modality, talk more in-depth about multimodal imaging and its benefits, and explore some of the newer topics emerging in the preclinical imaging world, including theranostics, contrast agent development, and many others. Please join us as we start this journey and continue to check back as we expand upon the basics introduced during this webinar.
Learning Objectives:
Understand convection-enhanced delivery and its implication for brain tumour treatment
Understand how gold nanoparticles can be used to construct radiation nanomedicine
Learn how to evaluate the safety, toxicity, and effectiveness of radiation nanomedicines
Overview:
Glioblastoma is a devastatingly aggressive type of brain tumour with a low median, and 5-year survival that has lacked new treatment options, in part due to the inability of therapeutic agents to cross the blood-brain barrier. Convection Enhanced Delivery (CED), a clinical neurosurgical strategy has been used to locoregionally deliver various therapeutic agents within the brain. Radiotherapeutic agents, such as 177Lu-labeled gold nanoparticles (177Lu-AuNP), hold promise for treatment of glioblastoma when administered by CED. Intratumoural injections of 177Lu-AuNP administered by CED was evaluated in an orthotopic xenograft mouse model of glioblastoma. SPECT/CT and biodistribution studies were used to evaluate the fate of the 177Lu-AuNP after injection. These results were used to estimate organ radiation absorbed doses. Normal tissue toxicity was evaluated to confirm the safety of the injections. Magnetic resonance imaging and bioluminescence imaging were used to monitor tumour growth after administration of 177Lu-AuNP, and median survival was estimated.
(February 16, 2023) Webinar: Intracerebral Transplantation of Autologous Bone...Scintica Instrumentation
Overview:
In this webinar, Max Myers presented his work on the use of autologous bone marrow-derived stem cells injected into the cortex of rats, following a stable stroke. Max also demonstrated its lab’s findings and talked about the Aspect Imaging M7 compact MRI system as it relates to its use in this project.
Key Points:
The critical use of stem cells in stroke research
Overcoming the blood-brain barrier via intracerebral injection of stem cells
The introduction of stem cells led to improved functional recovery following an ischemic stroke
How MRI can contribute to the understanding of treatments following stroke
Sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) uncoupling in skeletal muscle and mitochondrial uncoupling via uncoupling protein 1 (UCP1) in brown/beige adipose tissue are two primary mechanisms implicated in energy expenditure. Here, the effects of glycogen synthase kinase 3 (GSK3) inhibition via lithium chloride (LiCl) treatment on SERCA uncoupling in skeletal muscle and UCP1 expression in adipose were investigated. C2C12 and 3T3-L1 cells treated with LiCl had increased SERCA uncoupling and UCP1 protein levels, respectively, ultimately raising cellular respiration; however, this was only observed when LiCl treatment occurred throughout differentiation. In vivo, LiCl treatment (10 mg/kg/day) increased food intake in chow-fed and high-fat diet (HFD, 60% kcal) fed male mice without increasing body mass – a result attributed to elevated daily energy expenditure.
In soleus muscle, the lab determined LiCl treatment promoted SERCA uncoupling via increased expression of SERCA uncouplers, sarcolipin and/or neuronatin, under chow and HFD-fed conditions. They attribute these effects to the GSK3 inhibition observed with LiCl treatment as partial muscle specific GSK3 knockdown produced similar effects. In adipose, LiCl treatment inhibited GSK3 in inguinal WAT (iWAT) but not in brown adipose tissue under chow-fed conditions, which in turn led to an increase in UCP1 in iWAT and a beiging-like effect with a multilocular phenotype. The beiging-like effect was not observed, and increase in UCP1 when mice were fed a HFD, as LiCl could not overcome the ensuing overactivation of GSK3. Nonetheless, the study establishes novel regulatory links between GSK3 and SERCA uncoupling in muscle and GSK3 and UCP1 and beiging in iWAT.
Today, poultry meat is at the top of consumers’ lists as an affordable, nutritious, and healthy protein source. The modern meat-type chicken’s highly efficient growth rate and outstanding meat yield account for much of the poultry industry’s success, but many of today’s consumers care more than the price tag. As concerns for animal welfare, health, and sustainability grow along with the global population’s craving for animal protein, effectively addressing lameness incidence in meat birds is of key interest. A major obstacle in tackling lameness is the lack of understanding of the mechanistic underpinnings involved in a common cause of lameness, femur head necrosis (FHN), also called bacterial chondronecrosis with osteomyelitis (BCO).
The work of Dr. Alison Ramer’s lab has been to identify mechanisms by which bacteria induce the symptoms of FHN/BCO, bone attrition, inflammation, and infection through both in vivo and in vitro studies. This work has resulted in several key molecular pathways implicated in FHN/BCO, including DICER dysregulation and dsRNA accumulation, mitochondrial dysfunction, and autophagy dysregulation. In addition to identifying causative pathways, the lab has also sought to identify potential non-invasive biomarkers for the presence of FHN/BCO, which led to the identification of a unique cytokine/chemokine signature both in circulation and at the local bone level of affected birds. Their quest to better define and diagnose this consequential skeletal disease also means seeking new means of measuring and visualizing bone health in meat-type birds to feed the future healthily and sustainably.
In this webinar, Katie will discuss the role hypoxia plays in disease progression and treatment response, specifically in cancer. She will also dive into the various molecular imaging technologies that can be used to visualize and assess hypoxia in preclinical cancer models. Some modalities that will be covered include magnetic resonance imaging (MRI), positron emission tomography (PET), and optical imaging.
Topics to be covered:
What is hypoxia?
Is there a link between hypoxia and cancer?
What imaging modalities can be used to visualize hypoxia in vivo?
What are the advantages and limitations of each technique?
What are some applications of hypoxia imaging?
Hypoxia has been shown to influence many facets of cancer including tumor growth, treatment response, and metastatic potential. Thus, the ability to noninvasively visualize hypoxia in vivo may be critical to understanding the underlying tumor biology, guiding treatment plans, and determining prognosis in the clinic.
Many different modalities have been used for preclinical hypoxia imaging. While some techniques have been around for decades and have extensive data behind them, others are emerging technologies that aim to overcome existing limitations in the field. Choosing the right modality can be challenging and is dependent on experimental conditions including tumor model, animal strain, and the desired measurement, as each technique will target a different aspect of hypoxia. In this webinar, we will discuss some molecular imaging techniques that can be used to visualize and characterize tumor hypoxia including MRI, PET, optical, and PAI. We will compare the various options, discuss the advantages and limitations of each approach, and show some examples of how scientists are using these techniques within their research.
References
Rebecca A. D’Alonzo, Suki Gill, Pejman Rowshanfarzad, Synat Keam, Kelly M. MacKinnon, Alistair M. Cook & Martin A. Ebert (2021) In vivo noninvasive preclinical tumor hypoxia imaging methods: a review, International Journal of Radiation Biology, 97:5, 593-631, DOI: 10.1080/09553002.2021.1900943
(December 2, 2021) The Bench to Bedside Series: Preclinical Cancer Research w...Scintica Instrumentation
Overview:
The goal of this webinar will be to provide a high-level overview of the various stages of preclinical cancer research and discuss the role that innovative instrumentation can play in moving science forward.
To better understand how to treat and control cancer, researchers start by investigating the basics – the cells, molecules, and genes that make up the human body. This type of study, which is often referred to as basic or discovery research, aims to understand the underlying mechanisms contributing to cancer growth and spread. This knowledge is an essential starting point for developing future diagnostic tests and treatment strategies.
After finding an innovative idea that works in cells, researchers need to take their studies to the next level by employing animal models that have similar biology to humans. Animal models have helped scientists make some of the most important cancer discoveries over the years. Furthermore, preclinical imaging technologies allow researchers to perform longitudinal animal studies that are noninvasive leaving the underlying biology intact so that one can track changes throughout the entire disease process.
It was previously thought that the journey from bench to bedside was unidirectional, starting with discovery research and moving towards clinical trials. However, in the last decade, it has become crucial for basic scientists and clinicians to work together towards finding innovative solutions that will positively impact patient care.
After attending this webinar, we hope you will have a better understanding of the preclinical workflow needed to push an idea from bench to bedside as well as some of the key equipment that is needed along the way.
This webinar series will be hosted by Drs. Katie Parkins and Tyler Lalonde, both of which have extensive experience in translational research areas including oncology, neuroscience, molecular imaging, and drug development.
In this webinar we will discuss the following topics:
• Introduction To Cancer Research
• What does “Bench to Bedside” mean?
• In vitro characterization
• Rapid throughput screening
• Quantitative tools
• Moving towards translation
Overview:
Muscles are vital for everyday life, from every move we make to every beat of the heart. Conditions that lead to muscle wasting can drastically reduce our health and quality of life. This presentation will discuss the possibility of inhibiting an enzyme called glycogen synthase kinase 3 (GSK3) for the treatment/management of muscular dystrophy and spaceflight.
Without providing too much detail we will show our results with tideglusib treatment - a clinically advanced GSK3 inhibitor - on mdx mice. We will also discuss some of our ideas moving forward with spaceflight and how we plan on leveraging new infrastructure.
Objectives:
The importance of muscle health for overall health
Glycogen synthase kinase 3 and its role in regulating muscle size and composition
Calcium regulation in the heart
Muscular dystrophy
Spaceflight
Model Attribute Check Company Auto PropertyCeline George
In Odoo, the multi-company feature allows you to manage multiple companies within a single Odoo database instance. Each company can have its own configurations while still sharing common resources such as products, customers, and suppliers.
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
Embracing GenAI - A Strategic ImperativePeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
The Roman Empire A Historical Colossus.pdfkaushalkr1407
The Roman Empire, a vast and enduring power, stands as one of history's most remarkable civilizations, leaving an indelible imprint on the world. It emerged from the Roman Republic, transitioning into an imperial powerhouse under the leadership of Augustus Caesar in 27 BCE. This transformation marked the beginning of an era defined by unprecedented territorial expansion, architectural marvels, and profound cultural influence.
The empire's roots lie in the city of Rome, founded, according to legend, by Romulus in 753 BCE. Over centuries, Rome evolved from a small settlement to a formidable republic, characterized by a complex political system with elected officials and checks on power. However, internal strife, class conflicts, and military ambitions paved the way for the end of the Republic. Julius Caesar’s dictatorship and subsequent assassination in 44 BCE created a power vacuum, leading to a civil war. Octavian, later Augustus, emerged victorious, heralding the Roman Empire’s birth.
Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
The Roman Empire’s society was hierarchical, with a rigid class system. At the top were the patricians, wealthy elites who held significant political power. Below them were the plebeians, free citizens with limited political influence, and the vast numbers of slaves who formed the backbone of the economy. The family unit was central, governed by the paterfamilias, the male head who held absolute authority.
Culturally, the Romans were eclectic, absorbing and adapting elements from the civilizations they encountered, particularly the Greeks. Roman art, literature, and philosophy reflected this synthesis, creating a rich cultural tapestry. Latin, the Roman language, became the lingua franca of the Western world, influencing numerous modern languages.
Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
For more information, visit-www.vavaclasses.com
Acetabularia Information For Class 9 .docxvaibhavrinwa19
Acetabularia acetabulum is a single-celled green alga that in its vegetative state is morphologically differentiated into a basal rhizoid and an axially elongated stalk, which bears whorls of branching hairs. The single diploid nucleus resides in the rhizoid.
Instructions for Submissions thorugh G- Classroom.pptxJheel Barad
This presentation provides a briefing on how to upload submissions and documents in Google Classroom. It was prepared as part of an orientation for new Sainik School in-service teacher trainees. As a training officer, my goal is to ensure that you are comfortable and proficient with this essential tool for managing assignments and fostering student engagement.
Francesca Gottschalk - How can education support child empowerment.pptxEduSkills OECD
Francesca Gottschalk from the OECD’s Centre for Educational Research and Innovation presents at the Ask an Expert Webinar: How can education support child empowerment?
Francesca Gottschalk - How can education support child empowerment.pptx
Vascular Aging – New Lessons From a Novel "Knock-In" Mouse Model
1. Lakshmi Santhanam, Ph.D.
Associate Professor
Anesthesiology and Critical
Care Medicine
Scintica Instrumentation
Phone: +1 (519) 914 5495
sales@scintica.com
Vascular Aging – New
Lessons From a Novel
“Knock-In” Mouse Model
2. Vascular Aging – New Lessons
From a Novel “Knock-In” Mouse
Model
Lakshmi Santhanam, Ph.D.
Department of Anesthesiology & CCM
Johns Hopkins University School of Medicine
3. Objectives
• Mechanical perspective of vascular structure and function
• Understand the biophysical, biochemical, and cellular
hallmarks of vascular stiffening
• Tissue transglutaminase as a target in vascular aging
• Determine TG2’s mechanisms of action in the vasculature
• Understand cellular vs. matrix contributions to overall PWV in
mouse models.
4. Vascular Stiffness – Mechanical Perspective
The aorta as a non-linelarly viscoelastic material
Systole - Expansion
energy storage
Diastole – elastic recoil
unidirectional blood flow 𝑆𝑡𝑟𝑒𝑠𝑠; 𝜎 =
𝐹𝑜𝑟𝑐𝑒
𝐴𝑟𝑒𝑎
𝑆𝑡𝑟𝑎𝑖𝑛; 𝜆 =
𝐶ℎ𝑎𝑛𝑔𝑒 𝑖𝑛 𝑑𝑖𝑚𝑒𝑛𝑠𝑖𝑜𝑛
𝑂𝑟𝑖𝑔𝑖𝑛𝑎𝑙 𝑑𝑖𝑚𝑒𝑛𝑠𝑖𝑜𝑛
𝑆𝑡𝑟𝑎𝑖𝑛 𝑟𝑎𝑡𝑒 =
𝑑𝜆
𝑑𝑡
Loading – increasing stress
Unloading – decreasing stress
Einc depends on
Distending pressureWang et al (2016)
5. Load-bearing elements of the vascular wall
Elastic part: ECM
• Elastin fiber integrity
• Elastin/Collagen ratio
Elastin engages at low distension
Collagen engages at high distention
Physiologic range of pressure engages both
Viscous part: Incompletely understood
• Cell content
• VSMC tone
Shadwick (1995)
J. Exp. Biol 202:3305–3313
Wolinsky and Glagov (1964)
Circ Res. 14:400–413
6. Vascular Compliance – An Essential
Element of Overall Cardiovascular Health
Compliant Stiff
Windkessel
effect
Pulsatile flow input Steady flow
• Minimizes pump work needed
• Avoids cyclic loading/unloading of end organs –
protects from flow induced damage
• Optimal transport across capillaries
7. Pulse Wave Velocity – an Index of in
vivo Vascular Stiffness
Image from indusinstruments.com
𝑃𝑊𝑉
=
𝐸𝑖𝑛𝑐ℎ
𝐷𝜌
Where
Einc = incremental elastic modulus
h = wall thickness
D= lumen diameter
𝜌 = blood density
Moens Korteweg equation:
• Longitudinal studies
• Minimal discomfort to
mouse
• No damage to vasculature
can be used for ex vivo
measures
Doppler PWV:
Non-invasive PWV measurement
8. 20 40 60 80 100 120
3.0
3.5
4.0
4.5
5.0
MAP (mmHg)
PWV(m/s)
SNP infusion
PE infusion
L
Transducer 1
Transducer 2
Catheter
Catheter
Drug infusion
(PE to 120 mm Hg; SNP to 50 mm Hg)
ECG
d2P/dt2
PWV = L/Δt
Δt
Proximal wave
Distal wave
Einc = Incremental elastic modulus
h = wall thickness
r = radius of vessel
ρ = blood density
Moens – Korteweg Equation:
MAP dependence of Pulse Wave Velocity:
Simultaneous PWV-MAP Measurement (Invasive)
If HTN Is noted, PWV must be corrected for or
normalized against MAP
10. European Heart Journal, Volume 31, Issue 19, October 2010, Pages 2338–2350, EUROPEAN CONSORTIUM
Pulse Wave Velocity Increases with Age and Hypertension
N = 11,092 subjectsN = 1455 subjects; Normal PWV
Numbers at the tops of bars represent the overall percentage distribution of all subtypes of untreated hypertension
in that age group.
IDH (SBP <140 mm Hg and DBP ≥90 mm Hg) – Pharmacologics available
SDH (SBP ≥140 mm Hg and DBP ≥90 mm Hg) – IDH drugs + Lifestyle modifications
ISH (SBP ≥140 mm Hg and DBP <90 mm Hg) – NO PHARMACOLOGICAL AGENTS!
Franklin et al. Hypertension. 2001 Mar;37(3):869-74
11. PWV and Aging in Rodents
Mice Rats
Steppan et al, AJP Heart and Circ Physiol., 2019
Steppan et al, JAHA, 2014
12. PWV is a valuable predictor of cardiovascular risk
Ben Shlomo et al (2014); JACC 63:636-636
17,635
subjects
• aPWV predicts future cardiovascular events and mortality, even after accounting for
other established cardiovascular risk factors.
• Predictor of coronary heart disease and stroke after adjusting for traditional risk
factors
• Predictive value is stronger in younger versus older subjects – improved
classification particularly for younger individuals at intermediate risk
• Predictive value was not modified by hypertension, smoking, sex, diabetes, or
kidney disease.
• Improved the overall 10-year classification by 13%.
Lifestyle
Vs.
GeneticsAge
Diabetes
Hypertension
Obesity
Co-morbidities
Exercise
Diet
Stress
13. Endothelial Cells (EC)
Smooth Muscle Cells (VSMC)
Nitric Oxide (NO)
Blood pressure (cGMP)
Protein function (S-nitrosylation)
Protein secretion (“secretome”)
Gene expression
↑ Matrix Stiffness (↑collagen, ↑ elastin
fragmentation, ↑remodeling)
↑ Cell stiffness
Molecular Events
Structural Changes
↑ Vascular Stiffness
↑ VSMC motility, proliferation,
de-differentiation
Cell behavior
Vascular Function
Molecular/ Cellular Hallmarks of Vascular Stiffening
Aging
Endothelial dysfunction:
↓ NO Bioavailability
↓ Barrier function
Aberrant Cell-matrix
interaction/mechano-transduction
15. Tissue transglutaminase (TG2): A Matrix
Remodeling Enzyme
“Open”
“Closed”
GTP
Ca2+
https://pdb101.rcsb.org/motm/209
b-sandwich Catalytic core Barrel 1 Barrel 2 Full length TG2(4 domains)
Transamidation Reaction = Classical
Transglutaminase Function
• Member of transglutaminase family
• 1 of 8 catalytically active members
• >80% TG2 is cytosolic
• <20% at cell membrane/ ECM
• Secretion occurs via Golgi-independent pathway
• Activated in wound healing, fibrosis
Lysine Glutamine
Isopeptide
bond
16. Bakker et al, Circ Res, 2006
Eftekhari et al, J Vasc Res, 2007
Tissue Transglutaminase (TG2)
Background
So…Does TG2 play a role in
regulating stiffness in the
aorta?
TG2 mediates resistance
vessel remodeling
Low flow induced remodeling PE induced vasoconstriction
Purified TG2
+ CysNO
Lai TS et al, Biochemistry
40(16),
4904-4910 (2001)
17. TG2 secretion is NO dependent
= TG2 Red = Intracellular
Green = Extracellular
NO?
GSNO = NO donor
Cyst = TG inhibitor
L-NAME = eNOS inhibitor
DTT = reducing agent
Santhanam et al, Circ Res, 2010
18. Endothelial NO regulates TG2 secretion and activity
Santhanam et al, Circ Res, 2009
Santhanam et al, Chem Biol, 2011
Jandu et al, Amino Acids, 2011
Co-culture approach
HAEC = endothelial cells
HASMC = smooth muscle cells
L-NAME = NOS inhibitor
HAEC = human aortic endothelial cells
HASMC = human aortic SMC
IMR-90 = human Fibroblast
19. Hypothesis
Increased Central Vascular Stiffness
TG2
NO
SNO
Inhibits cross-linking function
eNOS
Hypothesis
ROSNO
eNOS
H2O2
O2-.
AGINGDecreased NO bioavailability
Increased ROS
Disrupt cell signaling
Impair vascular function
NOX enzymes
Mitochondrial enzymes
20. TG2-SNO decreases and transamidation activity increases with age
Decellularize
Homogenize tissue scaffold
Determine Matrix TG2 by
Western Blotting
= TG2
= VSMC
= EC
L-NAME
BASE
E-
YOUNG
OLD
O1 O2 Y1 Y2
Santhanam et al; Circ Res 2009
Jandu et al, Amino Acids, 2011
Cystamine - + - +
Young Old
BPA
Total TG2
RAT HUMAN
33-49 Years 62-101 Years
TG activity increases with age
in human aorta
21. TG2 is the primary vascular TGase
WT TG2-/-± L-NAME
20 mg/kg/day
4 weeks
Santhanam et al, Circ Res, 2010
-L682.777 + L682.777
80% TGase activity in the aorta is
TG2 derived
(Green = TG2 activity; blue = nucleus)
WT aorta becomes stiffer; but TG2 KO mice are
protected in L-NAME induced endothelial dysfunction.
L-NAME = NOS inhibitor
Doppler PWV Pressure
myography
L-NAME = eNOS inhibitor
Compliance = inverse of stiffness
22. 0.0 0.5 1.0 1.5 2.0
0
1000
2000
3000
Strain
Stress,mN/mm2
TG2-/-
WT
**
A) B)
TG2-/- mice: In vivo and ex vivo stiffness
TG2
GAPDH
WT
TG2-/-
Young mice
(Decellularized matrix)
Tensile testing
Decellularize
Steppan et al, JAHA; 2017 Feb 3;6(2):e004161.
Armstrong et al Acta Cir Bras. 2018 Nov;33(11):991-999.
C)
23. TG2-/- mouse aorta is more compliant in vitro but
not in vivo
D) E)
50 70 90 110 130 150
2.0
2.5
3.0
3.5
4.0
4.5
5.0
MAP (mmHg)
PWV,m/s
WT
TG2-/-
0.0 0.5 1.0 1.5 2.0
0
500
1000
1500
2000
Strain
Stress,mN/mm2
TG2-/-
WT
**
Intact vessels
VSMCs compensate for more compliant matrix to maintain baseline physiological stiffness
Steppan et al, JAHA; 2017 Feb 3;6(2):e004161.
24. b-sandwich Catalytic core Barrel 1 Barrel 2 Full length TG2 (4 domains)
What is the mechanism?
CYTOSOL
EXTRACELLULAR MATRIX
(Protein crosslinking – classic
TGase reaction)
CELL SURFACE
(Cell adhesion – Fibronectin,
Integrin, syndecan, binding)
INTRACELLULAR/ CELL
MEMBRANE (INNER)
(GTPase/ GTP binding)
= GTP
= Ca2+
1. Gundemir, Biochim. Biophys. Acta, 2012: 1823(2):406-19
2. Mehta K. Transglutaminases. 2015:215-228.
3. Kang et al, Biochem Biophys Res Commun. 2002 Apr
26;293(1):383-90
Ca2+
GTP
Closed
conformation
Open
conformation
TG2: A molecular “Swiss-army
knife”
• GTPase PLC VSMC tone
• TG2-adhesive function
Strength/number of focal
adhesions
• Traction force
• VSMC stiffness
4. Feng et al, Biochemistry. 1999 Feb 16;38(7):2224-32
5. Belkin and co-workers – TG2-integrin/Fn interaction
6. Griffin and co-workers – TG2- syndecan interaction
25. IP: Integrin β1
WB: TG2
WB: Integrin β1
WT
eNOSKO
WT eNOS KO
0
100
200
300
400
TG2/Integrin
(NormalizedtoWT)
*
(n = 5)
IgGcontrol
Jung et al, AJP Heart, 2013
Steppan et al, JAHA, 2017
What is the Mechanism?
Magnetic twisting cytometry
𝑆𝑡𝑖𝑓𝑓𝑛𝑒𝑠𝑠 ∝
1
𝐷𝑖𝑠𝑝𝑙𝑎𝑐𝑒𝑚𝑒𝑛𝑡
1. Tighter TG2 mediated
cell adhesion
2. Cell Stiffness
WT TG2-/-
0.0
0.2
0.4
0.6
0.8
1.0
CellStiffness(Pa/nm)
P=0.0019
26. Exogenous TG2 recovers passive stiffnessand
vasoreactivity in TG2-/- mice independent of crosslinking
function
L682.777 = Specific TG2 inhibitor
gpTG2 = purified guinea pig liver transglutaminase
rTG2 = recombinant TG2
27. TGM2-C277S mouse: A novel knock-in
mouse
sgRNA
Cas9 nickase
HDR Oligo
Fertilized
oocytes
Surrogate
mother
Screen to ID
F0 (Founders)
sgRNA Target C277 active site cysteine
HDR Oligo sequence bearing desired mutation
Santhanam Lab, unpublished results
30. Matrix remodeling and VSMCs are both
shareholders in vascular stiffening
• Aortic stiffness increases with age in WT mice
• TG2-/- mice are protected from aging associated stiffening
• TGM2-C277S mice Stiffer than TG2-/-, but marked protection vs. WT
A) B)
ECMTG2
Total TG2
GAPDH
Y O Y O
WT TGM2-C277S
3-6mo
>15mo
3-6mo
>15mo
3-6mo
>15mo
0
2
4
6
8
PWV(m/s)
WT
C277S
****
TG2-/-
****
****
**
**
Santhanam Lab, unpublished results
31. Matrix and VSMC both contribute to passive
stiffness
****
****
****
****
Santhanam Lab, unpublished results
32. Targeting TG2 crosslinking function interrupts
vascular stiffening but does not reverse it
± Cystamine (TG inhibitor)
40 mg/kg/day
3 months
Santhanam et al, Circ Res., 2010
Cystamine = TG inhibitor (non-specific)
34. Conclusions
• TG2 is an important target in aging-associated vascular
stiffening
• TG2 is regulated in the vasculature by an NO-dependent
mechanism
• The novel TGM2-C277S knock-in mouse shows matrix and
VSMC are both important shareholders in vascular stiffening
• VSMC-Matrix interaction/biomechanical input from the matrix
is an important determinant of VSMC tone/stiffness in vivo
• Targeting individual functions of TG2 can yield distinct changes
to overall stiffness by altering the VSMC stiffness and/or
VSMC-ECM crosstalk
• This model can be used to determine the specific role of TG2’s
crosslinking function in other diseases.
35. Acknowledgements
Dr. Dan Berkowitz
Dr. Daniel Nyhan
Dr. Steven An
Dr. Marc Halushka
Dr. Alberto Avolio
Dr. Alexey Belkin
Dr. Mark Butlin
Dr. Jochen Steppan
Dr. Sungmee Jung
Dr. Dinani Armstrong
Dr. Young Jun Oh
Funding:
NIH R01-HL105296
AHA BGIA2220181
ACCM StAAR awards
Dr. Simran Jandu
Dr. Alanah Webb
James Chen
Sean Melucci
Ivy Wang
Sandeep Jandu
Kavitha Nandakumar
Mentors/collaborators
Postdoctoral
Mentees/Fellows
Pre-doc Mentees/
Technicians
36. To ask a question, click the Q&A Button,
type your question and click send. Any
questions that are not addressed during
the live webinar will be answered
following the event.
Please indicate whom you want to
address your question.
Thank you for participating!
Q&A
SESSION:
Lakshmi Santhanam, Ph.D.
Associate Professor
Anesthesiology and Critical
Care Medicine