This study investigated whether perlecan Domain V (DV) and its C-terminal fragment LG3 can inhibit amyloid-β (Aβ) toxicity and restore angiogenic function in brain endothelial cells. The study found that:
1) Aβ reduced proliferation and tube formation of brain microvascular endothelial cells in vitro, while DV and LG3 blocked these effects of Aβ and restored proliferation and tube formation.
2) DV restored the ability of brain endothelial cells to form tube-like structures on Matrigel in the presence of Aβ, and this effect was dependent on the α5β1 integrin.
3) DV and LG3 may work through both the α5β1
This document describes a new bioinformatic procedure for identifying the genetic basis of physiological processes in non-model species. The procedure is illustrated by identifying genes involved in cold-induced angiogenesis in carp, a non-model species. The strategy uses ortholog prediction between carp, zebrafish and human to link carp DNA sequences to known angiogenic genes. Conditioned stepped reciprocal best hit BLAST analysis was found to be the most effective ortholog assignment method. Applying this strategy to carp gene expression data identified 135 candidate genes involved in cold-induced angiogenesis, 12 of which were examined in more detail.
Life is extremely precious and no one is guaranteed anything. Injuries, neurological disorders, and autoimmune disorders can make life extremely difficult. If a person has any of these conditions, he or she can be put on a transplant list to be given a second chance.
Stem cells are found in the umbilical cord of a newborn baby and they have been used to treat over 80 diseases. Banking your cord blood is vital because it will give your baby a perfect match for transplant if something should happen later. Your baby’s cord blood could be a possible match for you and for the rest of your family, too.
Our new eBook, the Latest Clinical Update for Stem Cell Therapies, is designed to show you how umbilical cord blood can treat nearly 20 different diseases and disorders. This is just a small insight into the importance of cord blood and why banking can save a family member’s life. With each informed person, more lives are saved.
Pascal J. Goldschmidt gave a presentation on designer genes and atherosclerosis. He discussed how vascular progenitor cell senescence can lead to plaque instability. Gene expression profiling was used to identify molecular signatures of arterial repair and different stages of atherosclerosis. Certain genes like LSAMP were found to predict left main coronary artery disease. Genomic techniques can now be used to predict atherosclerosis, coronary artery disease, and plaque vulnerability to help improve testing and prevention. Collaborative work was presented involving multiple departments at Duke University studying various aspects of atherosclerosis at the genetic and molecular level.
- Corazonin, AKH, and ACP are present in clusters of cells in the central nervous system of Rhodnius prolixus. AKH is also present in the corpus cardiacum.
- AKH increases lipid levels in the haemolymph, while corazonin increases heartbeat frequency. No effects were observed for ACP.
- The distribution and physiological roles of these neuropeptides in R. prolixus were mapped through immunohistochemistry and assays measuring lipid mobilization and heart contraction frequency.
Altered cholesterol intracellular trafficking contributes to the development of pathological hallmarks of sporadic Alzheimer's disease (AD). The presence of the APOE4 allele, the strongest genetic risk factor for sporadic AD, leads to impaired cholesterol recycling and accumulation of cholesterol in endolysosomes. This disrupts endolysosome function and contributes to increased amyloid-beta production, neurofibrillary tangle formation, and synaptic dysfunction—the key pathological hallmarks of AD. Targeting altered cholesterol homeostasis pathways may offer therapeutic strategies for treating or preventing sporadic AD, though cholesterol-lowering drugs like statins are not suitable due to potential worsening of endolysosome dysfunction.
There is promising research into developing vaccines against atherosclerosis. Studies have found that vaccination against oxidized LDL and certain ApoB-100 peptides can reduce the development of atherosclerosis in animal models by up to 60%. A recent study immunized ApoE null mice with ApoB-100 peptide sequences and found a 60% reduction in atherosclerotic plaque in the aorta. Further research is still needed but vaccination shows potential as a way to prevent atherosclerosis and vulnerable plaques in humans. International collaboration could help advance this area towards developing heart attack prevention through immune modulation.
Vaccination against atherosclerosis shows promise as an exciting new therapy. Studies in animal models have found that vaccination against oxidized LDL or specific ApoB-100 peptides can significantly reduce atherosclerosis. This study investigated immunization with novel ApoB-100 peptide sequences in ApoE null mice. It found a 60% reduction in atherosclerotic plaque in the aorta of immunized mice, along with increased collagen content in plaques. While more research is still needed, these results suggest that targeted vaccination against antigens involved in atherosclerosis, such as specific oxidized LDL peptides, could potentially be developed into a novel strategy for preventing heart attacks and strokes.
1) The presentation hypothesizes that chronic stress can cause bipolar disorder by upregulating the endocrine stress response and releasing high levels of glucocorticoids.
2) These glucocorticoids are absorbed by astrocytes in the prefrontal cortex and can lead to their degeneration over time.
3) Experiments in mice and cell cultures provide evidence that glucocorticoid exposure reduces astrocyte number and function in the prefrontal cortex and hippocampus.
This document describes a new bioinformatic procedure for identifying the genetic basis of physiological processes in non-model species. The procedure is illustrated by identifying genes involved in cold-induced angiogenesis in carp, a non-model species. The strategy uses ortholog prediction between carp, zebrafish and human to link carp DNA sequences to known angiogenic genes. Conditioned stepped reciprocal best hit BLAST analysis was found to be the most effective ortholog assignment method. Applying this strategy to carp gene expression data identified 135 candidate genes involved in cold-induced angiogenesis, 12 of which were examined in more detail.
Life is extremely precious and no one is guaranteed anything. Injuries, neurological disorders, and autoimmune disorders can make life extremely difficult. If a person has any of these conditions, he or she can be put on a transplant list to be given a second chance.
Stem cells are found in the umbilical cord of a newborn baby and they have been used to treat over 80 diseases. Banking your cord blood is vital because it will give your baby a perfect match for transplant if something should happen later. Your baby’s cord blood could be a possible match for you and for the rest of your family, too.
Our new eBook, the Latest Clinical Update for Stem Cell Therapies, is designed to show you how umbilical cord blood can treat nearly 20 different diseases and disorders. This is just a small insight into the importance of cord blood and why banking can save a family member’s life. With each informed person, more lives are saved.
Pascal J. Goldschmidt gave a presentation on designer genes and atherosclerosis. He discussed how vascular progenitor cell senescence can lead to plaque instability. Gene expression profiling was used to identify molecular signatures of arterial repair and different stages of atherosclerosis. Certain genes like LSAMP were found to predict left main coronary artery disease. Genomic techniques can now be used to predict atherosclerosis, coronary artery disease, and plaque vulnerability to help improve testing and prevention. Collaborative work was presented involving multiple departments at Duke University studying various aspects of atherosclerosis at the genetic and molecular level.
- Corazonin, AKH, and ACP are present in clusters of cells in the central nervous system of Rhodnius prolixus. AKH is also present in the corpus cardiacum.
- AKH increases lipid levels in the haemolymph, while corazonin increases heartbeat frequency. No effects were observed for ACP.
- The distribution and physiological roles of these neuropeptides in R. prolixus were mapped through immunohistochemistry and assays measuring lipid mobilization and heart contraction frequency.
Altered cholesterol intracellular trafficking contributes to the development of pathological hallmarks of sporadic Alzheimer's disease (AD). The presence of the APOE4 allele, the strongest genetic risk factor for sporadic AD, leads to impaired cholesterol recycling and accumulation of cholesterol in endolysosomes. This disrupts endolysosome function and contributes to increased amyloid-beta production, neurofibrillary tangle formation, and synaptic dysfunction—the key pathological hallmarks of AD. Targeting altered cholesterol homeostasis pathways may offer therapeutic strategies for treating or preventing sporadic AD, though cholesterol-lowering drugs like statins are not suitable due to potential worsening of endolysosome dysfunction.
There is promising research into developing vaccines against atherosclerosis. Studies have found that vaccination against oxidized LDL and certain ApoB-100 peptides can reduce the development of atherosclerosis in animal models by up to 60%. A recent study immunized ApoE null mice with ApoB-100 peptide sequences and found a 60% reduction in atherosclerotic plaque in the aorta. Further research is still needed but vaccination shows potential as a way to prevent atherosclerosis and vulnerable plaques in humans. International collaboration could help advance this area towards developing heart attack prevention through immune modulation.
Vaccination against atherosclerosis shows promise as an exciting new therapy. Studies in animal models have found that vaccination against oxidized LDL or specific ApoB-100 peptides can significantly reduce atherosclerosis. This study investigated immunization with novel ApoB-100 peptide sequences in ApoE null mice. It found a 60% reduction in atherosclerotic plaque in the aorta of immunized mice, along with increased collagen content in plaques. While more research is still needed, these results suggest that targeted vaccination against antigens involved in atherosclerosis, such as specific oxidized LDL peptides, could potentially be developed into a novel strategy for preventing heart attacks and strokes.
1) The presentation hypothesizes that chronic stress can cause bipolar disorder by upregulating the endocrine stress response and releasing high levels of glucocorticoids.
2) These glucocorticoids are absorbed by astrocytes in the prefrontal cortex and can lead to their degeneration over time.
3) Experiments in mice and cell cultures provide evidence that glucocorticoid exposure reduces astrocyte number and function in the prefrontal cortex and hippocampus.
Vitamin C has roles as both an antioxidant and in collagen production. A study found that vitamin C deficiency in mice compromised collagen deposition in atherosclerotic plaques, reducing collagen content. However, vitamin C levels did not affect plaque size or foam cell formation. While vitamin C may stabilize plaques by increasing collagen, more research is needed to determine if excess vitamin C can reverse atherosclerosis or if local vitamin C treatment could increase plaque stability. The study demonstrates vitamin C's role in collagen production for plaque stability but did not determine its dominant role in atherosclerosis.
Vitamin C has roles as both an antioxidant and in collagen production. A study found that vitamin C deficiency in mice compromised collagen deposition in atherosclerotic plaques, reducing collagen content. However, vitamin C levels did not affect plaque size or foam cell formation. While vitamin C may stabilize plaques by increasing collagen, more research is needed to determine if excess vitamin C can reverse atherosclerosis or if local vitamin C treatment could increase plaque stability. The study demonstrates vitamin C's role in collagen production for plaque stability but did not determine its dominant role in atherosclerosis.
CD33 is a genetic risk factor for late-onset Alzheimer's disease. Research found that CD33 inhibits the uptake of amyloid beta by microglia in the brain. Studies in mice and human brain tissue showed that deleting or inhibiting CD33 promotes the clearance of amyloid beta by microglia. This identifies a novel pathway by which the CD33 gene impacts amyloid accumulation in the aging brain and suggests targeting CD33 activity may help slow cognitive decline in Alzheimer's disease.
The document discusses complement activation and traumatic brain injury. It summarizes several studies that show:
1. Complement proteins like C3 and the membrane attack complex C5b-9 are activated in the border zone of brain contusions in rats and humans following traumatic brain injury.
2. There is local synthesis of complement proteins like C3, and upregulation of the regulator clusterin in response to complement attack on neurons and astroglia.
3. Complement activation occurs endogenously in the injured brain without circulating blood components, as shown in studies using brain slice cultures.
4. Genetic factors influence the inflammatory response and complement activation following TBI, with more pronounced immune reactivity linked
This study developed a new method for heat induced epitope retrieval (HIER) that allows staining of the cerebral vasculature in thick tissue samples. By using a class of closely related compounds including compound B in HIER, antibodies were able to bind to vascular proteins and stain the entire cerebral vasculature. Testing showed this method worked across species by staining canine and human tissue, and with multiple vascular antibodies. Further experiments revealed compounds A and C also allowed vascular staining with less tissue damage than compound B. This new HIER method utilizing these compounds provides a simple way to analyze the cerebral vasculature in disease models.
Resuscitation Science tips: The NYC Project Hypothermia rationale for phase IIEmergency Live
Resuscitation Science tips: The NYC Project Hypothermia
Intra-arrest induction of Therapeuitic Hypothermia via large-volume ice-cold saline infusion improves immediate outcomes for out-of-hospital cardiac arrest
The original source of this article is the AHA Journlas
http://circ.ahajournals.org/cgi/content/meeting_abstract/124/21_MeetingAbstracts/A2
Background: New York City Project Hypothermia is a collaborative effort involving the Fire Department of New York (FDNY), Greater New York Hospital Association, Health and Hospitals Corporation, the Regional Emergency Medical Advisory Committee, and the New York State Department of Health. As part of this effort, the FDNY implemented a pilot protocol in the New York City 9-1-1 System on August 1, 2010 that introduced the induction of therapeutic hypothermia during initial resuscitation efforts via large-volume ice-cold saline infusion.
Purpose: We sought to assess the effects of this protocol on immediate survival end-points following out-of-hospital cardiac arrest (OOHCA).
Methods: OOHCA data was analyzed for the following periods: August 1, 2009 - May 31, 2010 (historicalcontrol group) and August 1, 2010 - May 31, 2011 (study group). Except for the intra-arrest induction of hypothermia, no other aspect of the regional resuscitation protocols differed between the two periods. Standard Utstein definitions were utilized. Due to the large sample sizes, Chi-square analyses without Yates' correction were utilized, and a p <0.05 was considered significant.
Results: 5,582 resuscitations for nontraumatic adult cardiac arrests during the control period were compared to 4,727 resuscitations in the study period that included the intra-arrest induction of hypothermia. The groups did not differ with respect to age, response time, bystander witnessed status, or frequency of bystander CPR. Patients in the study period were less likely to be male (52.3% vs 54.6%, p = 0.019), less likely to be white (32.8% vs 35.1%, p = 0.013), and less likely to have an EMS-witnessed arrest (8.3% vs 9.5%, p=0.026). Return of spontaneous circulation (ROSC) and sustained ROSC were improved in the study group as compared to the control group: 31.7% vs 29.0% (p=0.003) and 24.1% vs 21.9% (p=0.0014), respectively.
The administration of large-volume, ice-cold saline for the intra-arrest initiation of therapeutic hypothermia improves immediate survival for out-of-hospital cardiac arrest.
Further work is required to assess the impact of this effect on long-term, neurologically intact survival and specific patient population for which this therapy may be of greatest benefit.
Special Thanks from the AHA to All of the Certified First Responder, Emergency Medical Technicians, and Paramedics of the FDNY and the New York City 9-1-1 System.
Để xem full tài liệu Xin vui long liên hệ page để được hỗ trợ
: https://www.facebook.com/thuvienluanvan01
HOẶC
https://www.facebook.com/garmentspace/
https://www.facebook.com/thuvienluanvan01
https://www.facebook.com/thuvienluanvan01
tai lieu tong hop, thu vien luan van, luan van tong hop, do an chuyen nganh
This document discusses new concepts in the treatment of pediatric traumatic brain injury. It covers several topics:
1) Age-dependent injury patterns and pathology occur following pediatric TBI, with infants more commonly experiencing diffuse injuries like swelling compared to focal injuries seen in older children.
2) Both immediate (primary) and delayed (secondary) injury can occur, with secondary injury involving an endogenous cascade leading to ongoing axonal injury and cell damage. Necrosis and apoptosis contribute to cell death.
3) Cerebral swelling is an important contributor to intracranial hypertension in children and may result from osmolar shifts, cellular edema, and blood-brain barrier breakdown. Traumatic axonal injury also commonly occurs.
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.
Gene polymorphisms in the renin-angiotensin-aldosterone system (RAAS) have been associated with differences in myocardial perfusion as measured by myocardial perfusion imaging (MPI). Specifically, the ACE I/D, REN C5312T, AGT M235T, AGT T174M, AT1R A1166C, and AT2R C3123A polymorphisms have been investigated for their relationship to MPI results. Studies have found that certain alleles or combinations of alleles of these RAAS gene polymorphisms are independent predictors of summed stress scores and summed difference scores on MPI and can influence myocardial blood flow.
Oakes et al 2017 TBK1 - a new player in ALS linking autophagy and neuroinflam...Maria Davies
TBK1 is a kinase involved in innate immunity and autophagy pathways. Recent human genetics studies have linked mutations in TBK1 to amyotrophic lateral sclerosis (ALS), a neurodegenerative disease affecting motor neurons. TBK1 plays a major role in autophagy by phosphorylating autophagy adaptor proteins, and several other ALS-linked genes are also involved in autophagy. Mutations in TBK1 may impair autophagy and contribute to the accumulation of protein aggregates, a hallmark of ALS pathology. The review discusses the role of TBK1 in autophagy and how disrupted autophagy may underlie the pathogenesis of ALS
Following ischemic stroke, activating cannabinoid receptor types 1 and 2 (C...Andreea-Diana Moisa
Following ischemic stroke, activating cannabinoid receptor types 1 and 2 (CB1 and CB2) using endogenous and synthetic agonists appears to have neuroprotective effects by reducing calcium influx, inhibiting glutamate release to normalize homeostasis, reducing oxidative stress, and restoring blood supply. CB1 activation enhances endocannabinoid signaling to promote neuronal maintenance and function, while CB2 activation protects against cerebral ischemia by inhibiting neutrophil recruitment and reducing infarct size.
This document is a request from a County Department of Family and Children Services to a medical provider to complete a physical examination for an employee or applicant. The employee or applicant would work as a Family Service Worker, whose responsibilities could include parenting, home management, personal care such as bathing and dressing, and transportation assistance for children, disabled individuals, and the elderly. The medical provider is asked to determine if the employee or applicant has the physical and medical capacity to perform these tasks on a full-time basis and note any restrictions.
Este discurso celebra os 20 anos da Universidade Politécnica e a graduação de 2015. Agradece a oportunidade dada aos estudantes de fazer parte da construção da instituição e contribuir para seu crescimento. Reconhece o trabalho do corpo docente e administrativo por levar a bom porto o percurso acadêmico dos estudantes com sucesso. Expressa orgulho pela universidade ser um exemplo de democracia no país.
Ever wondered what makes a really great product? Or what is the difference between good and great user experience? This presentation will give you some really cool answers covering those topics and also will encourage you to... make more mistakes. True story.
Este documento presenta los grupos y reglas para un torneo de pádel mixto. El torneo consta de dos grupos con parejas distribuidas. Cada victoria otorga 3 puntos mientras que cada derrota con un set ganado da 2 puntos y con dos sets perdidos da 1 punto. Las cuatro mejores parejas de cada grupo avanzan a la fase final de cuartos, semifinales y final.
Dr. Jai Singh is an Assistant Professor in the Department of India and World Literatures at The English and Foreign Languages University in Hyderabad. He received his MA from Kurukshetra University and his PhD from EFLU. His areas of specialization include contemporary theory, postcolonial theory and literature, Indian writing in English, feminism, and cultural studies. He has published papers in several international and national journals, presented at numerous conferences, and supervised students for their MA dissertations and ITP projects.
Cw 566 family services quarterly statistical reportscreaminc
This quarterly statistical report from the Georgia Department of Human Resources provides county-level data on family services programs and caseloads. It includes the number of adults and children served across programs like APS, CPS, and EMP. Staffing information is also presented, listing the number of state-funded family service workers, other support staff, and positions from other organizations that provide services.
An emergency intake form was completed for a child entering foster care, providing key medical and family information. The form documents the child's name, birthdate, physician contact details, any siblings or parents in the system, reasons for removal such as abuse or neglect, known allergies or medical issues, dietary needs or medications. It also notes any medical equipment or records sent with the child, their temporary placement, and contact details for the person completing the form.
Vitamin C has roles as both an antioxidant and in collagen production. A study found that vitamin C deficiency in mice compromised collagen deposition in atherosclerotic plaques, reducing collagen content. However, vitamin C levels did not affect plaque size or foam cell formation. While vitamin C may stabilize plaques by increasing collagen, more research is needed to determine if excess vitamin C can reverse atherosclerosis or if local vitamin C treatment could increase plaque stability. The study demonstrates vitamin C's role in collagen production for plaque stability but did not determine its dominant role in atherosclerosis.
Vitamin C has roles as both an antioxidant and in collagen production. A study found that vitamin C deficiency in mice compromised collagen deposition in atherosclerotic plaques, reducing collagen content. However, vitamin C levels did not affect plaque size or foam cell formation. While vitamin C may stabilize plaques by increasing collagen, more research is needed to determine if excess vitamin C can reverse atherosclerosis or if local vitamin C treatment could increase plaque stability. The study demonstrates vitamin C's role in collagen production for plaque stability but did not determine its dominant role in atherosclerosis.
CD33 is a genetic risk factor for late-onset Alzheimer's disease. Research found that CD33 inhibits the uptake of amyloid beta by microglia in the brain. Studies in mice and human brain tissue showed that deleting or inhibiting CD33 promotes the clearance of amyloid beta by microglia. This identifies a novel pathway by which the CD33 gene impacts amyloid accumulation in the aging brain and suggests targeting CD33 activity may help slow cognitive decline in Alzheimer's disease.
The document discusses complement activation and traumatic brain injury. It summarizes several studies that show:
1. Complement proteins like C3 and the membrane attack complex C5b-9 are activated in the border zone of brain contusions in rats and humans following traumatic brain injury.
2. There is local synthesis of complement proteins like C3, and upregulation of the regulator clusterin in response to complement attack on neurons and astroglia.
3. Complement activation occurs endogenously in the injured brain without circulating blood components, as shown in studies using brain slice cultures.
4. Genetic factors influence the inflammatory response and complement activation following TBI, with more pronounced immune reactivity linked
This study developed a new method for heat induced epitope retrieval (HIER) that allows staining of the cerebral vasculature in thick tissue samples. By using a class of closely related compounds including compound B in HIER, antibodies were able to bind to vascular proteins and stain the entire cerebral vasculature. Testing showed this method worked across species by staining canine and human tissue, and with multiple vascular antibodies. Further experiments revealed compounds A and C also allowed vascular staining with less tissue damage than compound B. This new HIER method utilizing these compounds provides a simple way to analyze the cerebral vasculature in disease models.
Resuscitation Science tips: The NYC Project Hypothermia rationale for phase IIEmergency Live
Resuscitation Science tips: The NYC Project Hypothermia
Intra-arrest induction of Therapeuitic Hypothermia via large-volume ice-cold saline infusion improves immediate outcomes for out-of-hospital cardiac arrest
The original source of this article is the AHA Journlas
http://circ.ahajournals.org/cgi/content/meeting_abstract/124/21_MeetingAbstracts/A2
Background: New York City Project Hypothermia is a collaborative effort involving the Fire Department of New York (FDNY), Greater New York Hospital Association, Health and Hospitals Corporation, the Regional Emergency Medical Advisory Committee, and the New York State Department of Health. As part of this effort, the FDNY implemented a pilot protocol in the New York City 9-1-1 System on August 1, 2010 that introduced the induction of therapeutic hypothermia during initial resuscitation efforts via large-volume ice-cold saline infusion.
Purpose: We sought to assess the effects of this protocol on immediate survival end-points following out-of-hospital cardiac arrest (OOHCA).
Methods: OOHCA data was analyzed for the following periods: August 1, 2009 - May 31, 2010 (historicalcontrol group) and August 1, 2010 - May 31, 2011 (study group). Except for the intra-arrest induction of hypothermia, no other aspect of the regional resuscitation protocols differed between the two periods. Standard Utstein definitions were utilized. Due to the large sample sizes, Chi-square analyses without Yates' correction were utilized, and a p <0.05 was considered significant.
Results: 5,582 resuscitations for nontraumatic adult cardiac arrests during the control period were compared to 4,727 resuscitations in the study period that included the intra-arrest induction of hypothermia. The groups did not differ with respect to age, response time, bystander witnessed status, or frequency of bystander CPR. Patients in the study period were less likely to be male (52.3% vs 54.6%, p = 0.019), less likely to be white (32.8% vs 35.1%, p = 0.013), and less likely to have an EMS-witnessed arrest (8.3% vs 9.5%, p=0.026). Return of spontaneous circulation (ROSC) and sustained ROSC were improved in the study group as compared to the control group: 31.7% vs 29.0% (p=0.003) and 24.1% vs 21.9% (p=0.0014), respectively.
The administration of large-volume, ice-cold saline for the intra-arrest initiation of therapeutic hypothermia improves immediate survival for out-of-hospital cardiac arrest.
Further work is required to assess the impact of this effect on long-term, neurologically intact survival and specific patient population for which this therapy may be of greatest benefit.
Special Thanks from the AHA to All of the Certified First Responder, Emergency Medical Technicians, and Paramedics of the FDNY and the New York City 9-1-1 System.
Để xem full tài liệu Xin vui long liên hệ page để được hỗ trợ
: https://www.facebook.com/thuvienluanvan01
HOẶC
https://www.facebook.com/garmentspace/
https://www.facebook.com/thuvienluanvan01
https://www.facebook.com/thuvienluanvan01
tai lieu tong hop, thu vien luan van, luan van tong hop, do an chuyen nganh
This document discusses new concepts in the treatment of pediatric traumatic brain injury. It covers several topics:
1) Age-dependent injury patterns and pathology occur following pediatric TBI, with infants more commonly experiencing diffuse injuries like swelling compared to focal injuries seen in older children.
2) Both immediate (primary) and delayed (secondary) injury can occur, with secondary injury involving an endogenous cascade leading to ongoing axonal injury and cell damage. Necrosis and apoptosis contribute to cell death.
3) Cerebral swelling is an important contributor to intracranial hypertension in children and may result from osmolar shifts, cellular edema, and blood-brain barrier breakdown. Traumatic axonal injury also commonly occurs.
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.
Gene polymorphisms in the renin-angiotensin-aldosterone system (RAAS) have been associated with differences in myocardial perfusion as measured by myocardial perfusion imaging (MPI). Specifically, the ACE I/D, REN C5312T, AGT M235T, AGT T174M, AT1R A1166C, and AT2R C3123A polymorphisms have been investigated for their relationship to MPI results. Studies have found that certain alleles or combinations of alleles of these RAAS gene polymorphisms are independent predictors of summed stress scores and summed difference scores on MPI and can influence myocardial blood flow.
Oakes et al 2017 TBK1 - a new player in ALS linking autophagy and neuroinflam...Maria Davies
TBK1 is a kinase involved in innate immunity and autophagy pathways. Recent human genetics studies have linked mutations in TBK1 to amyotrophic lateral sclerosis (ALS), a neurodegenerative disease affecting motor neurons. TBK1 plays a major role in autophagy by phosphorylating autophagy adaptor proteins, and several other ALS-linked genes are also involved in autophagy. Mutations in TBK1 may impair autophagy and contribute to the accumulation of protein aggregates, a hallmark of ALS pathology. The review discusses the role of TBK1 in autophagy and how disrupted autophagy may underlie the pathogenesis of ALS
Following ischemic stroke, activating cannabinoid receptor types 1 and 2 (C...Andreea-Diana Moisa
Following ischemic stroke, activating cannabinoid receptor types 1 and 2 (CB1 and CB2) using endogenous and synthetic agonists appears to have neuroprotective effects by reducing calcium influx, inhibiting glutamate release to normalize homeostasis, reducing oxidative stress, and restoring blood supply. CB1 activation enhances endocannabinoid signaling to promote neuronal maintenance and function, while CB2 activation protects against cerebral ischemia by inhibiting neutrophil recruitment and reducing infarct size.
This document is a request from a County Department of Family and Children Services to a medical provider to complete a physical examination for an employee or applicant. The employee or applicant would work as a Family Service Worker, whose responsibilities could include parenting, home management, personal care such as bathing and dressing, and transportation assistance for children, disabled individuals, and the elderly. The medical provider is asked to determine if the employee or applicant has the physical and medical capacity to perform these tasks on a full-time basis and note any restrictions.
Este discurso celebra os 20 anos da Universidade Politécnica e a graduação de 2015. Agradece a oportunidade dada aos estudantes de fazer parte da construção da instituição e contribuir para seu crescimento. Reconhece o trabalho do corpo docente e administrativo por levar a bom porto o percurso acadêmico dos estudantes com sucesso. Expressa orgulho pela universidade ser um exemplo de democracia no país.
Ever wondered what makes a really great product? Or what is the difference between good and great user experience? This presentation will give you some really cool answers covering those topics and also will encourage you to... make more mistakes. True story.
Este documento presenta los grupos y reglas para un torneo de pádel mixto. El torneo consta de dos grupos con parejas distribuidas. Cada victoria otorga 3 puntos mientras que cada derrota con un set ganado da 2 puntos y con dos sets perdidos da 1 punto. Las cuatro mejores parejas de cada grupo avanzan a la fase final de cuartos, semifinales y final.
Dr. Jai Singh is an Assistant Professor in the Department of India and World Literatures at The English and Foreign Languages University in Hyderabad. He received his MA from Kurukshetra University and his PhD from EFLU. His areas of specialization include contemporary theory, postcolonial theory and literature, Indian writing in English, feminism, and cultural studies. He has published papers in several international and national journals, presented at numerous conferences, and supervised students for their MA dissertations and ITP projects.
Cw 566 family services quarterly statistical reportscreaminc
This quarterly statistical report from the Georgia Department of Human Resources provides county-level data on family services programs and caseloads. It includes the number of adults and children served across programs like APS, CPS, and EMP. Staffing information is also presented, listing the number of state-funded family service workers, other support staff, and positions from other organizations that provide services.
An emergency intake form was completed for a child entering foster care, providing key medical and family information. The form documents the child's name, birthdate, physician contact details, any siblings or parents in the system, reasons for removal such as abuse or neglect, known allergies or medical issues, dietary needs or medications. It also notes any medical equipment or records sent with the child, their temporary placement, and contact details for the person completing the form.
This profile is for an experienced professional seeking an administrative role where they can utilize their operational knowledge and experience. They have over 10 years of experience in manufacturing, processing, and operations roles for companies like Electrolux, Randstad, Simos Solution, and Williams-Sonoma. Their experience includes assembling parts, training new employees, ensuring proper inventory flow, and managing staff. They aim to help an organization achieve its goals and take on more responsibility to enhance their skills.
Это предложение для настоящих поклонников кофе. Зачем покупать дорогие кофемашины, если можно взять в аренду для офиса или дома. О преимуществах такого предложения узнаете из презентации.
This document outlines the pricing and payment terms for engineering, survey, and non-destructive testing (NDT) services provided by Clever Marine Services Ltd. It lists hourly and daily rates for various roles like supervising engineers, electricians, and NDT technicians. It also specifies terms related to travel expenses, payment due dates, termination notice periods, and dispute resolution through arbitration in London. The rates are listed in both US dollars and Euros and are valid 24/7 for one year.
Sprzedaż produktów/usług premium nie jest taka trudna, gdy się wie, kto je kupuje.
Pisałam kiedyś na ten temat - możesz przeczytać tu: http://ewagadomska.pl/marketing/kto-kupuje-produkty-luksusowe-trzy-typy-klienta/.
Senior Sem. Final Project-Professional MaterialsMia Matthews
Resveratrol is a natural compound that may help reduce beta-amyloid plaques and prevent/treat Alzheimer's disease. It has shown to bind to amyloid proteins, changing their shape to non-toxic forms and decreasing their accumulation in the brain. Resveratrol also has antioxidant properties that can reduce oxidative stress associated with amyloid plaque formation. While studies in cells and animals have shown promise, more research is still needed to determine resveratrol's long-term safety and effectiveness in humans for treating Alzheimer's.
This document discusses the potential of enhancing sAPPα as a therapeutic strategy for Alzheimer's disease and other neurodegenerative diseases. It begins by providing background on Alzheimer's disease and current treatments. It then discusses APP processing and the role of sAPPα as a trophic factor that supports synaptic function. Mutations in APP, ADAM10, and genes involved in APP processing that alter risk for Alzheimer's and affect sAPPα production are also reviewed. The document concludes that enhancing sAPPα may be a promising therapeutic approach as it could both increase neuronal health and decrease amyloid beta production.
1. Alzheimer's disease is a major health concern that causes cognitive decline through β-amyloid accumulation and plaque formation in the brain.
2. Resveratrol has been shown to inhibit β-amyloid accumulation by binding to the proteins and changing their conformation to a non-toxic form, reducing plaque formation and neuronal damage.
3. While studies support resveratrol's potential as a therapeutic for Alzheimer's disease, long-term clinical trials in humans are still needed to confirm its safety and efficacy.
Amyloid diseases- Is there light at the end of the tunnel_Jonathon Burke
The document discusses amyloid diseases, particularly Alzheimer's disease. It provides background on the amyloid cascade hypothesis proposed in 1907 linking amyloid plaques to Alzheimer's. Recent research further supports this link, finding amyloid-beta accumulation correlates with cognitive decline. Studies show amyloid-beta has negative effects on synaptic activity and brain connectivity. Early detection of amyloid pathology could allow earlier treatment before full Alzheimer's symptoms develop.
This document summarizes a study examining the effects of high glucose and diabetes on cellular proteasome function in retinal and kidney cells. The key findings are:
1) Retinal endothelial cells exhibited significantly higher proteasome peptidase activity compared to pericytes and other cell types. High glucose treatment increased proteasome activity in endothelial cells but decreased it in pericytes.
2) High glucose treatment increased total levels of ubiquitinated proteins in retinal pericytes and endothelial cells, but not in photoreceptor cells. It also elevated levels of the PA28-a/-b proteasome regulatory subunits in pericytes and other cell types.
3) Retinas from diabetic mice showed
This document discusses neurodegenerative diseases and their treatment. It defines neurodegenerative diseases as conditions that primarily affect neurons in the brain, causing problems with movement or mental functioning as neurons are damaged and die. The causes include a buildup of toxic proteins and mitochondrial dysfunction, generally resulting in programmed cell death of neurons. Stroke is also discussed, defined as occurring when a blood vessel blockage or rupture in the brain causes part of the brain to be deprived of oxygen. The two main types of stroke and their symptoms are outlined.
A New Perspective On Alzheimer S Disease As A Brain Expression Of A Complex M...Tony Lisko
This document discusses Alzheimer's disease as a brain expression of a complex metabolic disorder. It summarizes that AD is characterized by abnormalities at systemic, histological, macromolecular and biochemical levels beyond amyloid plaques and neurofibrillary tangles. It proposes that cerebral hypoperfusion and metabolic stress from factors like atherosclerosis, infections and insulin resistance can trigger AD pathology by inducing oxidative damage and impairing brain energy metabolism. Specifically, it suggests that arginine and branched chain amino acid metabolism disturbances contribute to AD pathogenesis and that maintaining adequate levels of these nutrients may have therapeutic potential.
ABSTRACT- Coronary artery disease (CAD) is suspected as a leading cause of mortality in developed countries. Due
to cholesterol and fat deposit plaque is forming into the inner walls of the arteries of the heart, which leads to narrowing
of blood vessels of heart and reduce the blood flow rate into heart. Proprotein convertase subtilisin-like kexin type 9
(PCSK9) is one of the candidate gene that regulate lipoprotein retention pathway of CAD development. It is a newly
discovered serine protease that plays a key role in LDL-C homeostasis by mediating LDL receptor (LDLR). The LDL
receptor is breakdown through a post transcriptional mechanism and induces the production of very low-density
lipoprotein in the fasting state. The aim of this study was to investigate the frequency of single nucleotide
polymorphism (SNP) of PCSK9 gene of 155 CAD patients and 102 ages matched healthy controls. Serum lipids
including total cholesterol (TC), triglycerides (TG), HDL, LDL, and VLDL were analyzed. PCR-RFLP analysis was
carried out to genotype regions carrying Eam 1104I restriction site in the PCSK9. Gene considering significant
difference in serum TC, TG, HDL-C, LDL-C and VLDL-C levels (P<0.001, <0.0001) of patients and control samples.
In CAD patients, G allele frequency is less than A allele frequency. G allele is responsible for decreasing the
LDL: HDL ratio which shows evidence in having its protecting effect on the occurrence of CAD in West Bengal Population.
Key-words- CAD, PCSK9, SNP, Eam1104I, Polymorphism, West Bengal population
The document discusses protein aggregation diseases, known as amyloidoses, which occur when certain proteins abnormally aggregate within or surrounding cells. These diseases range from systemic amyloidoses that can affect any part of the body to those that affect a single organ. Many neurodegenerative diseases are also protein aggregation disorders. The formation of protein aggregates depends on factors like protein concentration, interactions with other proteins, and the cellular environment. A better understanding of what influences the equilibrium between soluble and insoluble protein states is important for developing therapies for these diseases.
Caffeine, Through Adenosine A3 Receptor-Mediated Actions, Suppresses Amyloid-...Mahmoud Lotfy Soliman
1. Caffeine prevents the internalization of LDL cholesterol in neurons by blocking adenosine A3 receptors (A3Rs). Caffeine decreased neuronal internalization of DiI-labeled LDL cholesterol in a concentration-dependent manner.
2. Blocking A3Rs with a specific antagonist or knocking down A3Rs with siRNA mimicked caffeine's effects in decreasing LDL cholesterol internalization, while activating A3Rs increased internalization.
3. By blocking A3R-mediated internalization of LDL cholesterol, caffeine decreases amyloid beta (Aβ) generation from amyloid precursor protein (APP), which is increased by LDL cholesterol internalization through enhanced amyloidogenic processing of APP in endosomes.
This document provides an introduction to angiogenesis and anti-angiogenic therapy for brain tumors. It discusses how angiogenesis is important for tumor growth and how numerous growth factors and molecules regulate this process. Hypoxia within tumors can induce factors like VEGF that promote angiogenesis. The document outlines several pro-angiogenic growth factors and their receptors that are involved in glioma angiogenesis, including VEGF, FGF, HGF, angiopoietins and their roles. It also discusses endogenous inhibitors of angiogenesis like angiostatin and endostatin that may have therapeutic potential.
Konstantin Ravvin: Immune Therapy Targeting Tauopathies in Alzheimer's DiseaseKonstantin Ravvin
Konstantin Ravvin: Tauopathies are diseases related to abnormal filamentous inclusions of microtubule associated protein tau commonly found across a spectrum of neurodegenerative disorders. The dissociation of monomeric tau protein from microtubule bindings sites results in the formation of aggregates with a high affinity for self-assembly. Aggregates propagate interneuronally in a prion-like fashion, resulting in widespread neuronal damage, cognitive decline and cell death. tau’s co-association with amyloid-beta in Alzheimer’s diseases makes it a potential target for therapeutic intervention. More recently, immunotherapy in passive and active forms has been shown to ameliorate amyloid load and substantially rescue cognitive impairment. Antibody-mediated tau aggregation and clearance of tau aggregates has been described in previous studies though none have evaluated the effectiveness of antibodies at substochiometric concentrations sufficient to simulate the exclusivity of the blood brain barrier. We found that substochoichiometric concentrations of antibody binding to the C-terminus of tau exacerbated aggregation as measured by ThT
fluorescence. Fibril seeds formed in the presence of antibody decreased seeding efficiency relative to non-antibody treated seeds, and end products of antibody treated seeds were
substantially more structured relative to their unseeded and normally seeded counterparts, suggesting the potential formation of novel “strains”. Antibody in the presence of normal
seeds and monomeric tau lowered seeding efficiency in a concentration dependent manner, indicating a therapeutically optimal concentration of antibody adequate enough to
decrease deplete seeding efficiency without overtly intensifying monomeric aggregation.
This document summarizes a study investigating the neuroprotective effects of resveratrol in an Alzheimer's disease model. The study has two parts: 1) Improving resveratrol's bioavailability in vivo by increasing its availability at the target tissue through grafting of cells that promote resveratrol metabolite liberation. Different doses will be administered orally and intravenously to mice with and without these cells. 2) Examining resveratrol's effects on oxidative stress and inflammation in vitro in the presence and absence of a SIRT1 inhibitor to determine if these effects are SIRT1-independent. PC12 cells will be treated with beta-amyloid to induce toxicity with and without res
This document discusses Alzheimer's disease (AD), including its definition, etiology, risk factors, pathophysiology, clinical symptoms, diagnosis, and treatment. Some key points include:
- AD is the most common cause of dementia and is characterized by cognitive and behavioral impairment. While the exact cause is unknown, risk factors include age, family history, and genetics such as the APOE E4 allele.
- Pathologically, AD is defined by amyloid plaques and neurofibrillary tangles in the brain. It results from the death of brain cells, affecting processes like memory, thinking, and behavior.
- Diagnosis involves assessing symptoms, ruling out other conditions through tests, and structural imaging of the brain
Higher levels of the protein brain-derived neurotrophic factor (BDNF) may protect against Alzheimer's disease and dementia. BDNF targets cortical brain cells, preventing their death and improving learning and memory. Studies have also found that higher serum BDNF levels can protect against future occurrence of dementia and Alzheimer's. Future research may examine using BDNF levels to predict Alzheimer's risk or giving BDNF supplements to older adults to prevent or reduce symptoms.
This presentation illustrates the various pathways of development of AD ,including the recent molecular pathways , and their implication in early diagnosis and therapy .
This document summarizes a study on developing a novel protein interaction platform to study neurodegeneration. The study cultured neuronal stem cells to form neurospheres, which were used as a model system. Lentiviral transfection was optimized to introduce target genes stably into the neurospheres. The goal was to analyze protein interactions of Alzheimer's disease proteins by co-immunoprecipitation of neurosphere cultures expressing tagged target proteins. This model aims to further the understanding of molecular mechanisms in neurodegenerative disorders like Alzheimer's disease.
This document summarizes the role of the transcription factor nuclear factor-kappa B (NF-κB) in the major genetic and environmental risk factors for Alzheimer's disease (AD). It discusses how NF-κB interacts with genes implicated in AD risk, such as APP, APOE, and genes involved in immunity. It reviews evidence from neuronal cell lines, invertebrate models like fruit flies, and vertebrate models like mice that genetic and environmental factors associated with aging increase NF-κB activity and are linked to AD pathology. The document suggests NF-κB may be a key regulator integrating genetic and environmental risk factors and could potentially be targeted for disease prevention.
1) Alzheimer's disease is characterized by progressive cognitive decline associated with the propagation of amyloid-beta and tau protein fibrils in the brain, resulting in neuronal death.
2) The cholinergic hypothesis proposes that depleted levels of acetylcholine in the brain, particularly in areas responsible for memory and cognition, contribute to Alzheimer's symptoms. However, acetylcholine decline may be a symptom rather than a cause of neurodegeneration.
3) The amyloid cascade hypothesis posits that amyloid-beta plaque formation triggers tau protein tangle formation, driving the progression of Alzheimer's pathology and neurodegeneration.
Konstantin Ravvin (Sackler Journal of Medicine): The Biochemical Foundations...Konstantin Ravvin
1) Alzheimer's disease is characterized by the progressive spread of amyloid beta and tau protein fibrils in the brain, resulting in neuronal death and cognitive decline.
2) Early theories proposed that a decline in acetylcholine levels played a role in Alzheimer's, but it is now thought to be a symptom rather than a cause.
3) The amyloid cascade hypothesis posits that amyloid beta plaque formation triggers tau protein tangles, which together drive neurodegeneration. However, the relationship between amyloid beta and tau remains unclear.
2. INTRODUCTION
Alzheimer’s disease (AD) is the most common cause of dementia among the elderly with
symptoms including confusion, progressive loss of memory, and depression [1]. While the
neuropathological hallmarks of AD include extracellular deposition of amyloid-β (Aβ) in
the cortex and hippocampus and the intraneuronal accumulation of neurofibrillary tangles,
the deposition of Aβ on cerebral blood vessels is also a prominent component in AD [2],
resulting in neurovascular dysfunction and/or cerebral amyloid angiopathy [3]. A link
between AD and vascular disease has been suggested by many studies showing that
hypertension, atherosclerosis, stroke, diabetes mellitus, smoking, and high cholesterol are
common risk factors [4, 5]. Thus, neurovascular dysfunction is likely to be an important
component of dementia and AD.
While increases in Aβ production can contribute to AD pathogenesis, AD has been proposed
to be a “retention” disease caused by inefficient clearance of Aβ from the brain by the
blood-brain barrier (BBB) [3]. In healthy individuals, Aβ is cleared from the brain by
transport across the BBB via receptor-mediated transcytosis. If Aβ clearance pathways are
disrupted, soluble Aβ can accumulate [6] and promote the formation of neurotoxic Aβ
forms, resulting in neuronal and endothelial cell damage and death [2]. In traumatic brain
injury, a known predisposing factor for AD [7], disruption of the BBB is thought to result in
the higher risk of aberrant Aβ accumulation [8]. Therefore, functional endothelial cells are
critical to maintaining BBB integrity and regulating the BBB permeability for many solutes,
preserving brain homeostasis. Disruption of endothelial integrity enhances Aβ toxicity to
endothelial cells, disrupts Aβ clearance from the brain and contributes to the pathogenesis of
AD [9]. Additionally, Aβ can bind integrin receptors. Specifically, Aβ1-40 binds to cells
expressing the alpha2beta1 (α2β1) integrin, resulting in both Aβ deposition and
initialization of a neurotoxic signaling cascade [10]. Furthermore, the alpha5beta1 (α5β1)
integrin can mediate cell adhesion to Aβ1-40 and promote cellular internalization and
degradation of Aβ, resulting in an increase in the clearance of Aβ, a reduction in the
formation of an insoluble Aβ matrix, and a decrease of Aβ toxicity to cells [11].
The AD affected brain has an increase in proangiogenic factors such as vascular endothelial
growth factor (VEGF) [12]. This suggests that possible compensatory mechanisms exist to
counteract damaged vasculature in the brain. However, the low density of blood vessels seen
in AD suggests that this reparative angiogenic mechanism fails [13]. Aβ has been shown to
be anti-angiogenic in vitro and in vivo and thus its presence in the AD brain likely
contributes to the lack of new blood vessel formation [13]. Aβ also binds growth factor
receptors such as VEGFR2 thus blocking the angiogenic activity of VEGF on endothelial
cells [14]. Additionally, VEGF has been shown to co-aggregate with Aβ in plaques seen in
the AD brain [15], decreasing available VEGF to promote angiogenesis in the brain.
Therefore, Aβ appears to act as an antagonist for the proangiogenic factors in the brain,
opposing their effects on angiogenesis and maintenance of the cerebrovasculature [13].
Therefore, finding a way to re-establish the angiogenic system and promote blood vessel
formation could be an important component of successful AD treatment.
Parham et al. Page 2
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
3. The C-terminal portion of the extracellular matrix proteoglycan perlecan, termed Domain V
(DV), promotes brain-derived endothelial cell proliferation and is proangiogenic in the brain
in rodent stroke models [16]. It increases the production and release of VEGF in brain
endothelial cells via the α5β1 integrin and is neuroprotective [16]. DV consists of three
laminin-globular-like repeats (LG domains) separated by two type-1 epidermal growth
factor-like repeats. The C-terminal LG domain, LG3, has been shown to have many of the
functional activities of full-length DV [17]. LG3 is physically smaller than DV (26 kDa
versus 85 kDa, respectively) and thus, may be more tractable as a therapeutic molecule as it
is easier to produce and less susceptible to degradation.
α5β1 integrin is expressed in many cell types, including brain microvascular endothelial
cells, the latter during brain development and in circumstances such as brain injury, where it
helps promote angio-genesis and revascularization [18]. DV promotes angiogenesis through
the α5β1 integrin in brain microvascular endothelial cells and generally increases
endothelial cell health and function via expression and secretion of VEGF [19]. We
hypothesized that DV and LG3 could decrease Aβ-induced endothelial cell death and
prevent its antiangiogenic properties. In this study, we show that human DV and LG3 rescue
mouse brain endothelial cells from Aβ-induced cytotoxicity in vitro and that DV restores
Aβ-disrupted angiogenesis in an α5β1 integrin dependent manner. Additionally, we
demonstrate that both human and mouse LG3 are active in mouse brain endothelial cells.
MATERIALS AND METHODS
Cell culture
In vitro experiments were performed using the mouse cerebrovascular endothelial cell line,
MBEC. The MBEC cells were kindly provided by Dr. Jane Welsh (Texas A&M University)
and were isolated from the C57BL/6 strain of mice [16, 20]. The cells (passage 11–17) were
grown in Iscove’s Modified Dulbecco’s Medium (IMDM, Invitrogen, CA), 10% FBS, 1%
antibiotic/antimycotic (Cellgro), and 1% L-glutamine (Cellgro) on flasks coated with
porcine-derived gelatin (Sigma Aldrich).
α5 integrin knockdown
Cells were prepared as described in Lee et al. [16]. Briefly, MBECs were treated with α5
siRNA (Mission siRNA, Sigma-Aldrich) containing media with Lipofectamine 2000
(Invitrogen) and selected for resistance to puromycin to develop a stable cell line. α5
knockdown (75%) was previously confirmed by α5 quantitative PCR and by α5 western
blot analysis [16]. Cells were grown on flasks coated with porcine-derived gelatin (Sigma
Aldrich) in the media described for parental MBEC supplemented with 2 μg/ml puromycin.
DV and LG3 protein production
Human DV (hDV) and human LG3 (hLG3) were produced and purified from 293-EBNA
cells as previously published [16, 21]. In some experiments, hDV was heat inactivated for
15 min at 90°C. Mouse LG3 (mLG3) was cloned with a 3′ Histidine tag into the vector
pCep-pu (provided by Maurizio Mongiat, Center for Cancer Research, Aviano, Italy) using
the following primers: Mouse LG3 5′ Nhe I #2 5′-
Parham et al. Page 3
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
4. AGGCTAGCGTATGGGGTAGTGGAGTCAGACTG-GCAC-3′ and Mouse LG3 3′ Xho
I #2 5′-AGCTCGAGCATGATGATGATGATGATGTGAGG-
GGCAGGGCCGTGTGTTGG-3′. Mouse LG3 was amplified from full-length mouse DV
using Pfx50 (Invitrogen, Grand Island, NY). 293-EBNA cells were transfected via
Lipofectamine 2000 (Invitrogen) and stable cells were selected by growing the transfected
cells in basal medium supplemented with 0.05 μg puromycin. After transfection, conditioned
media from the transfected 293 EBNA cells was collected and mLG3 was purified using Ni-
NTA agarose beads (Qiagen, Valencia, CA). Eluted fractions containing mLG3 were
dialyzed against PBS and the purity was confirmed via SDS-PAGE stained with Brilliant
Blue G Colloidal (Sigma, St. Louis, MO) and western blot using an anti-6X His antibody
(Genetex, Irvine, CA). Human DV, hLG3, and mLG3 were used at 300 nM unless otherwise
indicated.
Aβ25-35 preparation
Human Aβ25-35 and the reverse peptide Aβ35-25 were purchased from California Peptide
Research, Inc. (Napa, CA) and were resuspended in sterile distilled H2O (Invitrogen,
Carlsbad, CA) to a final concentration of 1 mM. They were incubated for 3–4 days at 37°C
to allow the peptide to aggregate to its toxic form. Aggregated Aβ was stored at −80°C.
Capillary tube-like structure formation on a Matrigel matrix (in vitro angiogenesis assay)
MBECs in IMDM/1% FBS (1 × 105 per well) were treated in suspension for 20 min at
37°C/5% CO2 with hDV (300 nM), Aβ35-25 (25 μM) alone (reverse peptide), Aβ25-35 (25
μM) alone, Aβ25-35 plus hDV or IMDM/1% medium alone (control) and then added to
Matrigel (Becton Dickenson, Franklin Lakes, NJ) coated wells for 3–6 h also at 37°C/5%
CO2. The assay was stopped with 4% paraformaldehyde. Adobe Photoshop CS 5.0 for
Apple Macintosh was used to quantify pictures (three randomly selected microscopic fields
per well) for tube formation and calculating the average tube pixels per high power field as
previously published [16]. Results were expressed as a percentage of the control (arbitrarily
set to 100%) with error bars representing standard deviation. After the assay was stopped,
non-adherent cells in the media were harvested via centrifugation and quantified (percent
difference from control) with a hemocytometer using trypan blue stain to distinguish live
from dead cells.
Proliferation assay
MBECs were plated in 96-well tissue culture treated plates (1500 cells per well, triplicate
wells) in IMDM/1% FBS immediately after being exposed to hDV, hLG3, or mLG3 with or
without Aβ25-35 (25 μM) or media alone (control). Additionally, the cells were tested for
their response to an Aβ25-35 titration at 10, 25, or 50 μM. On day 3, the cells were treated
with alamar blue (AbD Serotec, Raleigh, NC) for 2–6 h followed by measurement with a
fluorescent plate reader, excitation at 560 nm and emission at 590 nm. The results were
expressed as percent change from the absorbance of the control (arbitrarily set at 100%).
Parham et al. Page 4
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
5. Statistical analysis
All experiments were repeated three independent times. The proliferation assay was
performed in triplicate per condition and the Matrigel experiments were performed in
duplicate per repeat. Individual experimental values (from repeated measures) were
averaged and used in subsequent statistical analyses. Statistical significance (p < 0.05) was
determined by Student’s t test. Error bars represent standard deviation unless otherwise
stated.
RESULTS
hDV and LG3 promote proliferation and overcome Aβ toxicity in mouse brain endothelial
cells
Cell proliferation is a required step during angiogenesis and DV has previously been
reported to increase proliferation in MBECs [16]. However, LG3 has not been evaluated for
this ability in these cells. Therefore, we tested the ability of hDV, hLG3, and mLG3 to
promote proliferation of MBECs (Fig. 1A). Cells were plated immediately after being
exposed to the test conditions and incubated in the wells for 72 h prior to alamar blue
analysis. hDV, but not heat inactivated hDV, promoted proliferation of MBECs. hLG3
significantly increased MBEC proliferation at 600 nM and 900 nM but not at the lower
concentration of 300 nM (Fig. 1A). Likewise, mLG3 was also unable to significantly
increase proliferation in the MBECs when at the lower concentration of 300 nM.
We next tested the capability of hDV, hLG3, and mLG3 to rescue Aβ-mediated endothelial
toxicity. MBECs were plated in the presence of Aβ25-35 ± DV, hLG3, or mLG3, and the
ability of the cells to proliferate was measured as described above. When the cells were
treated with Aβ25-35 only, MBECs showed decreased viability in a dose-dependent fashion
(Fig. 1B). The reverse peptide Aβ35-25 did not have any measurable effect in this assay.
Interestingly, hDV and mLG3 significantly rescued the MBECs from Aβ25-35 toxicity but
hLG3 did not.
DV rescues Aβ25-35 -induced inhibition of capillary tube-like formation by MBECs on a
Matrigel matrix
We next performed in vitro capillary “tubulogenesis” assays to determine if DV was able to
rescue MBEC from Aβ25-35-induced inhibition of the formation of capillary tube-like
structure formation on Matrigel (a growth factor enriched substance that promotes and
supports such tube-like structure) (Fig. 2). MBECs were seeded on Matrigel and treated with
media only, DV alone, Aβ25-35, DV plus Aβ25-35, or the reverse peptide Aβ35-25. We first
verified that DV increases the formation of tube-like structures in the MBECs as previously
published [16]. Aβ25-35 significantly inhibited tube formation while DV overcame Aβ25-35
inhibitive effects by promoting tube-like formation. The reverse peptide Aβ35-25 had no
toxic effects on the cells.
Reduction of capillary tube-like formation by Aβ25-35 is not due to cell death
During progression of the Matrigel assay, we noted that MBEC that were not part of the
tube-like structures were floating in the Aβ treated wells. To investigate if this was due to
Parham et al. Page 5
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
6. cell death, media containing the non-adhered cells was collected after assay progression was
stopped and cells were counted in the presence of trypan blue. Interestingly, upon
examination of the collected non-adhered cells, less than 1% were non-viable from all of the
treatment conditions, including the Aβ25-35 wells. MBEC treated with media only or DV did
not have many non-adhered cells and quantitation revealed that more than 90% of the cells
seeded were able to form tube-like structures. However, at 25μM Aβ25-35, 49% of the
original MBECs seeded were alive yet unable to adhere to form tubes (Fig. 3). Moreover
when DV was added along with Aβ25-35 at 25μM, it overcame Aβ25-35’s inhibition of cell
adhesion to the Matrigel matrix, showing a 35% decrease when compared to the 25 μM
Aβ25-35 alone wells.
α5β1 mediates rescue of capillary tube-like formation by DV
As stated earlier, DV induces VEGF secretion via the α5β1 integrin receptor [16]. If DV is
mediating rescue of the Aβ-induced reduction of capillary tube-like formation via the α5β1
integrin in the MBEC, we reasoned that DV should not rescue when the α5 integrin was
knocked down. α5 integrin- knockdown MBECs (MBEC α5 KD) that express 25% of
normal α5β1 integrin levels [16] were tested to determine if DV could restore their ability to
form tube-like structures in the presence of Aβ25-35 in the Matrigel assay (Fig. 4).
Surprisingly, DV was able to increase tube-like formation in the MBEC α5 KD cells by
72% suggesting that less than complete knockdown of α5β1 integrin was insufficient to
block DV tube promoting effects but also raises the possibility that other factors additionally
contribute in these MBECs. However, in contrast to wild-type MBECs, DV was unable to
rescue Aβ-induced reduction in tube formation.
DISCUSSION
In this study, we demonstrate that Aβ25-35 reduces proliferation of a mouse brain
microvascular endothelial cell line in vitro. Furthermore, we demonstrate that perlecan DV,
the 85 kDa C-terminal protein fragment of perlecan (itself a prominent heparan sulfate
proteoglycan component of the vascular basement membrane in the brain and throughout the
body), and mLG3 (the C-terminal LG domain of DV) blocks these Aβ25-35 effects on
proliferation. Interestingly, hLG3 required a 2-fold higher concentration than DV to increase
proliferation of MBECs. Mouse LG3 was more effective than hLG3 in blocking Aβ25-35
proliferation effects on MBECs but not more so in increasing MBEC proliferation by itself
without Aβ25-35. Mouse LG3 possesses a single glycosaminoglycan attachment site [22, 23]
not present in hLG3, potentially explaining the species specificity of the interaction.
Importantly, to the best of our knowledge, this is the first study that has examined the
function of mouse recombinant LG3.
In this study, we performed Matrigel assays to determine the ability of DV to overcome the
deleterious effects of Aβ25-35 on capillary tube-like formation. The formation of these
structures is thought to mimic the process by which endothelial cells form capillaries in vivo
[24]. We show that Aβ25-35 inhibits mouse brain microvascular endothelial cells from
forming a capillary network on Matrigel. It has been reported that Aβ1-40 has similar effects
on human brain-derived endothelial cells [13]. Furthermore, DV is able to restore the ability
Parham et al. Page 6
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
7. of MBECs to form tube-like structures in the presence of Aβ25-35. To our surprise, Aβ25-35
did not reduce these biological functions by inducing cell death as would be expected from a
cytotoxic agent. Angiogenesis is a tightly regulated process involving endothelial cell
proliferation, migration, attachment, recruitment of supporting cells, and the formation of
tubes [25]. Growth factors such as VEGF initiate angiogenesis and growth factor receptors
and integrins are crucial for attachment. In our study, there is a clear correlation between the
extent of tube formation by MBECs on Matrigel and cell adhesion to the matrix.
Additionally, the fact that cells treated with Aβ25-35 were alive but unable to form tubes
suggests that Aβ25-35 may cause the downregulation of a cell surface molecule critical for
adhesion or modify other events required for cell adhesion. We hypothesize that Aβ25-35,
like Aβ1-40, may bind to the integrins required for the cells to bind to Matrigel such as α5β1
and/or α2β1 and thereby inhibit their adhesion to Matrigel.
When α5β1 integrin expression was reduced by 75% in MBECs, DV was no longer able to
restore the reduction in tube-formation caused by Aβ25-35, demonstrating the importance of
the α5β1 integrin in mediating this activity of DV. Our lab has previously shown that α5
integrin knockdown in MBECs impaired their migratory capabilities when DV was the
migratory stimulus [19] and that the CRRETAWAC peptide (a peptide specific for the
binding pocket of the α5β1 integrin [26]) inhibited DV-induced capillary morphogenesis in
brain endothelial cells [16]. However, while blockade of α5β1 integrin could block DV-
induced capillary morphogenesis, the present study suggests that incomplete (75%)-α5
knockdown could not. While this is likely due to the residual α5β1 integrin still present on
the endothelial cells in the context of a strong angiogenesis promoting environment
(Matrigel is enriched in angiogenesis promoting growth factors), the possibility that other
factors could contribute to DV’s proangiogenic effect in this assay cannot be excluded and
merits further study. Nevertheless, α5β1 clearly plays an important, if not exclusive, role in
mediating DV’s activity on brain microvascular endothelial cells schematized in Fig. 5.
It is known that Aβ directly interacts with VEGFR2, blocking the ability of VEGF to bind to
VEGFR2 [14], the primary receptor for transducing angiogenic signals, preventing brain
microvascular endothelial cells from forming tube-like structures on Matrigel. Aβ disrupts
VEGF/VEGFR2 signaling by antagonizing VEGF-induced Akt phosphorylation and Tyr996
phosphorylation of VEGFR2 [14] (Fig. 5A). Previous studies have shown that exogenous
VEGF application can dose-dependently reverse the anti-angiogenic effect of Aβ [14],
VEGF levels in the brain are closely related to cognitive function [27–30] and
intraperitoneal injection of VEGF improved the cognitive impairment of AD mice [31].
Therefore, to reverse the deleterious effects of Aβ on angiogenesis, it is critical to disrupt
Aβ interaction with VEGFR2. Importantly, DV has been shown to directly bind VEGFR2
[32] and increase tube-like structure formation in brain endothelial cells by increasing their
VEGFR2 expression and VEGF secretion via the α5β1 integrin [16]. Additionally,
increasing VEGFR2 receptor and VEGF expression creates a proangiogenic positive
feedback loop. DV also binds α5β1 and when the α5β1 integrin is reduced, DV is unable to
induce VEGF secretion and thus is unable to rescue the reduction of Aβ in the formation of
tube-like structures. DV signals through α5β1 in MBECs, activating the Akt/ERK mitogen
activated pathway [19] and similarly, VEGF/VEGFR2 can signal through the same pathway,
inducing the expression of VEGF [33]. The blockade of ERK-dependent pathways upon
Parham et al. Page 7
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
8. decreased α5 integrin expression blunts DV-induced proliferation in brain endothelial cells
[19]. These proliferative signals induced by DV in endothelial cells might either overcome
the Aβ toxicity or interfere with Aβ-induced endothelial cell death. Furthermore, DV might
prevent Aβ from binding to VEGFR2 by directly binding to VEGFR2 itself and/or increase
the VEGF amount to a level where it can outcompete with Aβ for receptor binding and
subsequently induce an angiogenic signal.
Endothelial cells from the brains of AD transgenic mice are impaired in forming tube-like
structures on Matrigel, suggesting altered angiogenic properties in these mice [34].
Additionally, multiple in vitro and in vivo assays have shown that angiogenesis is inhibited
by soluble Aβ peptides. Finally, Aβ can inhibit the vascularization of xenografted human
cancer cells and oppose VEGF-induced angiogenesis in vivo [14]. Thus, the anti-angiogenic
properties of Aβ are well characterized and present a problem in the AD brain.
Understanding the molecular mechanisms behind the anti-angiogenic activity of Aβ and
discovering novel ways to overcome it will help us better evaluate the importance of
angiogenesis to the AD process and in particular the contribution of the cerebrovasculature
to the pathogenesis of AD. To this end, DV, a proteolytically cleaved C-terminal protein
fragment of the major vascular basement membrane component perlecan, and thereby
intimately associated with cerebrovasculature, may serve as part of the brain’s natural Aβ
defense mechanisms, inhibiting its deleterious effects on endothelial cell viability and
angiogene-sis when present. This concept is supported by the observation that the normally
ubiquitously expressed (around brain blood vessels) perlecan is conspicuously absent in
amyloid-laden brain blood vessels in human AD and cerebral amyloid angiopathy patients,
but present in amyloid-free “normal” brain blood vessels, suggesting that a lack of perlecan
(and its DV portion) could contribute to Aβ cerebrovascular pathology [35]. Our study
suggests that DV is beneficial to brain endothelial cells exposed to Aβ25-35 in vitro in an
integrin-mediated fashion. Future studies with other forms of Aβ, specifically Aβ1-40 and
Aβ1-42, as well as those involving in vivo models of AD and cerebral amyloid angiopathy,
may further support DV as a potential novel anti-amyloid therapy.
Acknowledgments
We would like to thank Dr. Michael Kahle (Texas A&M Health Science Center) and Dr. Aileen Marcelo
(University of Kentucky) for helpful comments during the preparation of this manuscript. Financial and material
support for this research was provided by the NIH R01NS065842-01A01 grant and the Ted Nash Long Life
Foundation Award.
References
1. Thies W, Bleiler L. 2011 Alzheimer’s disease facts and figures. Alzheimers Dement. 2011; 7:208–
244. [PubMed: 21414557]
2. Bell RD, Zlokovic BV. Neurovascular mechanisms and blood-brain barrier disorder in Alzheimer’s
disease. Acta Neuropathol. 2009; 118:103–113. [PubMed: 19319544]
3. Deane R, Zlokovic BV. Role of the blood-brain barrier in the pathogenesis of Alzheimer’s disease.
Curr Alzheimer Res. 2007; 4:191–197. [PubMed: 17430246]
4. Gorelick PB. Risk factors for vascular dementia and Alzheimer disease. Stroke. 2004; 35:2620–
2622. [PubMed: 15375299]
Parham et al. Page 8
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
9. 5. Rocchi A, Orsucci D, Tognoni G, Ceravolo R, Siciliano G. The role of vascular factors in late-onset
sporadic Alzheimer’s disease. Genetic and molecular aspects. Curr Alzheimer Res. 2009; 6:224–
237. [PubMed: 19519304]
6. Deane R, Sagare A, Zlokovic BV. The role of the cell surface LRP and soluble LRP in blood-brain
barrier Abeta clearance in Alzheimer’s disease. Curr Pharm Des. 2008; 14:1601–1605. [PubMed:
18673201]
7. Giunta B, Obregon D, Velisetty R, Sanberg PR, Borlongan CV, Tan J. The immunology of
traumatic brain injury: A prime target for Alzheimer’s disease prevention. J Neuroin-flammation.
2012; 9:185.
8. Pop V, Sorensen DW, Kamper JE, Ajao DO, Murphy MP, Head E, Hartman RE, Badaut J. Early
brain injury alters the blood-brain barrier phenotype in parallel with beta-amyloid and cognitive
changes in adulthood. J Cereb Blood Flow Metab. 2013; 33:205–214. [PubMed: 23149553]
9. Rizzo MT, Leaver HA. Brain endothelial cell death: Modes, signaling pathways, and relevance to
neural development, homeostasis, and disease. Mol Neurobiol. 2010; 42:52–63. [PubMed:
20407845]
10. Wright S, Malinin NL, Powell KA, Yednock T, Rydel RE, Griswold-Prenner I. Alpha2beta1 and
alphaVbeta1 integrin signaling pathways mediate amyloid-beta-induced neurotoxicity. Neurobiol
Aging. 2007; 28:226–237. [PubMed: 16448724]
11. Matter ML, Zhang Z, Nordstedt C, Ruoslahti E. The alpha5beta1 integrin mediates elimination of
amyloid-beta peptide and protects against apoptosis. J Cell Biol. 1998; 141:1019–1030. [PubMed:
9585419]
12. Paris D, Ganey N, Banasiak M, Laporte V, Patel N, Mullan M, Murphy SF, Yee GT, Bachmeier C,
Ganey C, Beaulieu-Abdelahad D, Mathura VS, Brem S. Impaired orthotopic glioma growth and
vascularization in transgenic mouse models of Alzheimer’s disease. J Neurosci. 2010; 30:11251–
11258. [PubMed: 20739545]
13. Paris D, Townsend K, Quadros A, Humphrey J, Sun J, Brem S, Wotoczek-Obadia M, DelleDonne
A, Patel N, Obregon DF, Crescentini R, Abdullah L, Coppola D, Rojiani AM, Crawford F, Sebti
SM, Mullan M. Inhibition of angiogenesis by Abeta peptides. Angiogenesis. 2004; 7:75–85.
[PubMed: 15302999]
14. Patel NS, Mathura VS, Bachmeier C, Beaulieu-Abdelahad D, Laporte V, Weeks O, Mullan M,
Paris D. Alzheimer’s beta-amyloid peptide blocks vascular endothelial growth factor mediated
signaling via direct interaction with VEGFR-2. J Neurochem. 2010; 112:66–76. [PubMed:
19818105]
15. Yang SP, Bae DG, Kang HJ, Gwag BJ, Gho YS, Chae CB. Co-accumulation of vascular
endothelial growth factor with beta-amyloid in the brain of patients with Alzheimer’s disease.
Neurobiol Aging. 2004; 25:283–290. [PubMed: 15123332]
16. Lee B, Clarke D, Al Ahmad A, Kahle M, Parham C, Auckland L, Shaw C, Fidanboylu M, Orr
AW, Ogunshola O, Fertala A, Thomas SA, Bix GJ. Perlecan domain V is neuroprotective and
proangiogenic following ischemic stroke in rodents. J Clin Invest. 2011; 121:3005–3023.
[PubMed: 21747167]
17. Bix G, Fu J, Gonzalez EM, Macro L, Barker A, Campbell S, Zutter MM, Santoro SA, Kim JK,
Hook M, Reed CC, Iozzo RV. Endorepellin causes endothelial cell disassembly of actin
cytoskeleton and focal adhesions through alpha2beta1 integrin. J Cell Biol. 2004; 166:97–109.
[PubMed: 15240572]
18. del Zoppo GJ, Milner R. Integrin-matrix interactions in the cerebral microvasculature. Arterioscler
Thromb Vasc Biol. 2006; 26:1966–1975. [PubMed: 16778120]
19. Clarke DN, Al Ahmad A, Lee B, Parham C, Auckland L, Fertala A, Kahle M, Shaw CS, Roberts J,
Bix GJ. Perlecan Domain V induces VEGf secretion in brain endothelial cells through integrin
alpha5beta1 and ERK-dependent signaling pathways. PLoS One. 2012; 7:e45257. [PubMed:
23028886]
20. Fuller A, Yahikozawa H, So EY, Dal Canto M, Koh CS, Welsh CJ, Kim BS. Castration of male
C57L/J mice increases susceptibility and estrogen treatment restores resistance to Theiler’s virus-
induced demyelinating disease. J Neurosci Res. 2007; 85:871–881. [PubMed: 17253641]
Parham et al. Page 9
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
10. 21. Wright S, Parham C, Lee B, Clarke D, Auckland L, Johnston J, Lawrence AL, Dickeson SK,
Santoro SA, Griswold-Prenner I, Bix G. Perlecan domain V inhibits alpha2 integrin-mediated
amyloid-beta neurotoxicity. Neurobiol Aging. 2012; 33:1379–1388. [PubMed: 21126803]
22. Friedrich MV, Gohring W, Morgelin M, Brancaccio A, David G, Timpl R. Structural basis of
glycosaminogly-can modification and of heterotypic interactions of perlecan domain V. J Mol
Biol. 1999; 294:259–270. [PubMed: 10556044]
23. Friedrich MV, Schneider M, Timpl R, Baumgartner S. Perlecan domain V of Drosophila
melanogaster. Sequence, recombinant analysis and tissue expression. Eur J Biochem. 2000;
267:3149–3159. [PubMed: 10824099]
24. Kubota Y, Kleinman HK, Martin GR, Lawley TJ. Role of laminin and basement membrane in the
morphological differentiation of human endothelial cells into capillary-like structures. J Cell Biol.
1988; 107:1589–1598. [PubMed: 3049626]
25. Lechertier T, Hodivala-Dilke K. Focal adhesion kinase and tumour angiogenesis. J Pathol. 2012;
226:404–412. [PubMed: 21984450]
26. Mould AP, Garratt AN, Puzon-McLaughlin W, Takada Y, Humphries MJ. Regulation of integrin
function: Evidence that bivalent-cation-induced conformational changes lead to the unmasking of
ligand-binding sites within integrin alpha5 beta1. Biochem J. 1998; 331(Pt 3):821–828. [PubMed:
9560310]
27. Wang Y, Galvan V, Gorostiza O, Ataie M, Jin K, Green-berg DA. Vascular endothelial growth
factor improves recovery of sensorimotor and cognitive deficits after focal cerebral ischemia in the
rat. Brain Res. 2006; 1115:186–193. [PubMed: 16928361]
28. Pati S, Orsi SA, Moore AN, Dash PK. Intra-hippocampal administration of the VEGF receptor
blocker PTK787/ZK222584 impairs long-term memory. Brain Res. 2009; 1256:85–91. [PubMed:
19100245]
29. Cao L, Jiao X, Zuzga DS, Liu Y, Fong DM, Young D, During MJ. VEGF links hippocampal
activity with neurogenesis, learning and memory. Nat Genet. 2004; 36:827–835. [PubMed:
15258583]
30. Plaschke K, Staub J, Ernst E, Marti HH. VEGF over-expression improves mice cognitive abilities
after unilateral common carotid artery occlusion. Exp Neurol. 2008; 214:285–292. [PubMed:
18822285]
31. Wang P, Xie ZH, Guo YJ, Zhao CP, Jiang H, Song Y, Zhu ZY, Lai C, Xu SL, Bi JZ. VEGF-
induced angiogenesis ameliorates the memory impairment in APP transgenic mouse model of
Alzheimer’s disease. Biochem Biophys Res Commun. 2011; 411:620–626. [PubMed: 21771586]
32. Goyal A, Pal N, Concannon M, Paul M, Doran M, Poluzzi C, Sekiguchi K, Whitelock JM, Neill T,
Iozzo RV. Endorepellin, the angiostatic module of perlecan, interacts with both the alpha2beta1
integrin and vascular endothelial growth factor receptor 2 (VEGFR2): A dual receptor antagonism.
J Biol Chem. 2011; 286:25947–25962. [PubMed: 21596751]
33. Claesson-Welsh L, Welsh M. VEGFA and tumour angiogenesis. J Intern Med. 2013; 273:114–127.
[PubMed: 23216836]
34. Paris D, Patel N, DelleDonne A, Quadros A, Smeed R, Mullan M. Impaired angiogenesis in a
transgenic mouse model of cerebral amyloidosis. Neurosci Lett. 2004; 366:80–85. [PubMed:
15265595]
35. van Horssen J, Otte-Holler I, David G, Maat-Schieman ML, van den Heuvel LP, Wesseling P, de
Waal RM, Verbeek MM. Heparan sulfate proteoglycan expression in cerebrovascular amyloid
beta deposits in Alzheimer’s disease and hereditary cerebral hemorrhage with amyloidosis (Dutch)
brains. Acta Neuropathol. 2001; 102:604–614. [PubMed: 11761721]
Parham et al. Page 10
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
11. Fig. 1.
DV and LG3 enhance brain endothelial cell proliferation and block Aβ25-35 toxicity. A) Quantification (% of control arbitrarily
set at 100%) of alamar blue measured proliferation of MBECs after 72 h in vitro treated as labeled. *p < 0.05 as compared to
control. H.i. hDV = heat-inactivated hDV. B) Quantification (as in A) with added Aβ25-35 ± DV or LG3 treatments as labeled
demonstrating Aβ25-35 dose dependent toxicity that was rescued by human DV and mLG3, but not hLG3 (all at 300 nM). *p <
0.05 as compared to 25 μM Aβ25-35.
Parham et al. Page 11
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
12. Fig. 2.
DV restores angiogenic capabilities to Aβ25-35-treated MBECs- No treatment control (A), DV treatment (B), Aβ35-25 reverse
peptide control (C), Aβ25-35 only (D), DV plus Aβ25-35 (E), and quantitation of three independent experiments (F). Calculations
were based on the average tube pixels per high power field (as described in the methods section) from three randomly selected
microscopic fields per well. Values are expressed as percent of control (no treatment) arbitrarily set to 100%. *p < 0.05 as
compared to indicated treatments.
Parham et al. Page 12
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
13. Fig. 3.
Reduction of capillary tube-like formation by Aβ25-35 is not due to cell death. Quantification of non-adhered cells from Matrigel
tube assay, treated as labeled. *p < 0.05 as compared to control.
Parham et al. Page 13
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
14. Fig. 4.
α5β1 mediates rescue of capillary tube-like formation by DV. Quantification (% of control, arbitrarily set at 100%) of tube
formation of MBEC α5 knockdown cells (75% knockdown) on Matrigel treated as indicated. *p < 0.05 as compared to control.
Parham et al. Page 14
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript
15. Fig. 5.
Schematic of proposed model. A) During AD pathogenesis, Aβ directly interacts with VEGFR2, blocking the ability of VEGF
to bind to VEGFR2, disrupting signaling. The complete Aβ/VEGFR2 pathway is currently unknown. B) DV directly binds both
VEGFR2 and the α5β1 integrin, increasing tube-like structure formation in brain endothelial cells by increasing VEGF secretion
and VEGFR2 expression, creating a proangiogenic positive feedback loop. When α5 expression is reduced, DV is unable to
induce VEGF secretion and thus is unable to rescue Aβ’s reduction in the formation of tube-like structures. DV signals through
α5β1 in MBECs to activate the Akt/ERK mitogen activated pathway. VEGF/VEGFR2 can signal through the same pathway,
inducing the expression of VEGF. We propose that DV prevents Aβ from binding to VEGFR2 by directly binding to VEGFR2
itself and/or by increasing the VEGF amount to a level where it can outcompete with Aβ for receptor binding and subsequently
induce an angiogenic signal.
Parham et al. Page 15
J Alzheimers Dis. Author manuscript; available in PMC 2015 January 01.
NIH-PAAuthorManuscriptNIH-PAAuthorManuscriptNIH-PAAuthorManuscript