The document examines the effects of the TACE inhibitor BMS-561392 on APP processing both in vitro and in vivo. In cell cultures expressing APP, BMS-561392 reduced secretion of sAPPα without increasing Aβ production. Conversely, a BACE inhibitor decreased sAPPβ and Aβ without affecting sAPPα. In vivo infusion of BMS-561392 into mouse brains decreased sAPPα levels but did not significantly change steady-state Aβ levels. The findings suggest that under normal conditions, BACE and TACE do not compete for APP as a substrate, though they share localization in the trans-Golgi network. Inhibition of TACE may therefore reduce TNFα levels in
Genetic and epigenetic biomarkers for therapeutic monitoring in neurological ...Pranav Sopory
Seminar held on Genetic and epigenetic biomarkers for therapeutic monitoring in neurological disorders.
Multiple Sclerosis, Alzheimer's Disease and Parkinson's Disease.
Biomarkers and epigenetic explained. New epigenetic drug targets.
1) Rats treated with 3-nitropropionic acid (3-NP), a model of Huntington's disease, exhibited weight loss, gait abnormalities, and striatal lesions.
2) The study found a dose-dependent reduction in complex-I activity in the cerebral cortex of 3-NP treated rats, as measured spectrophotometrically and by blue native-polyacrylamide gel electrophoresis.
3) Succinate driven State 3 respiration was significantly inhibited both in vivo and in isolated mitochondria from the cortex of 3-NP treated rats, suggesting complex-I dysfunction in addition to inhibition of complex-II and succinate dehydrogenase activity contributes to cortico-striatal lesions in this model
Screening models for Parkinson & multiple sclerosisHimanshu Yadav
1. The document discusses various in vivo and in vitro models used to study Parkinson's disease (PD). In vivo models include antagonism of tremorine/oxotremorine-induced parkinsonism in mice, reserpine antagonism in mice, the MPTP model in monkeys, and circling behavior in nigrostriatal lesioned rats. In vitro methods include experiments using rat striatal slices and measuring dopamine stimulated adenyl cyclase activity.
2. The experimental autoimmune encephalomyelitis (EAE) model is described as the most widely used animal model for multiple sclerosis (MS). EAE is induced by immunizing mice with myelin antigens, causing an inflammatory de
By: Edward M. Barbieri and James Shorter
TDP-43 forms cytoplasmic aggregates in degenerating neurons of frontotemporal dementia (FTD) patients.
Laferrière et al. now establish that TDP-43 assemblies from distinct FTD subtypes have different structures,
neurotoxicities, and seeding activities, which correlate with FTD severity. Thus, distinct pathological TDP-43
assemblies akin to prion strains might underpin distinct FTD subtypes.
This study investigated the role of the complement receptor C5ar1 in epilepsy models. The researchers found that C5ar1 was upregulated in mouse models of status epilepticus. Treatment with the C5ar1 antagonist PMX53 reduced seizures in acute and chronic seizure models. PMX53's anticonvulsant effects were mediated by C5ar1, as they were not seen in C5ar1-deficient mice or with an inactive analogue. Inhibition of C5ar1 during status epilepticus lessened seizures, protected neurons from degeneration, and reduced mortality. The study suggests blocking C5ar1 activation may be a novel target for developing new anti-epileptic drugs.
1) Human growth hormone (HGH) treatment increased survival motor neuron (SMN) protein levels in human neuronal cells in a dose-dependent manner by activating the STAT5 signaling pathway.
2) Systemic administration of HGH to spinal muscular atrophy (SMA) mouse models induced higher levels of SMN protein in the brain and spinal cord.
3) HGH treatment improved disease phenotypes and increased survival in two severe SMA mouse models, confirming that activating the STAT5 pathway may be a promising therapeutic strategy for SMA.
The document discusses various strategies for overcoming resistance to endocrine therapy in advanced breast cancer, including targeting downstream signaling pathways like PI3K/Akt/mTOR that are often aberrantly activated in endocrine resistant disease. It reviews clinical trials demonstrating improved progression-free survival when adding drugs like everolimus or CDK4/6 inhibitors to hormonal therapy. The goal is to inhibit compensatory survival pathways that become relied upon by cancer cells to drive continued growth in the face of endocrine therapy.
Genetic and epigenetic biomarkers for therapeutic monitoring in neurological ...Pranav Sopory
Seminar held on Genetic and epigenetic biomarkers for therapeutic monitoring in neurological disorders.
Multiple Sclerosis, Alzheimer's Disease and Parkinson's Disease.
Biomarkers and epigenetic explained. New epigenetic drug targets.
1) Rats treated with 3-nitropropionic acid (3-NP), a model of Huntington's disease, exhibited weight loss, gait abnormalities, and striatal lesions.
2) The study found a dose-dependent reduction in complex-I activity in the cerebral cortex of 3-NP treated rats, as measured spectrophotometrically and by blue native-polyacrylamide gel electrophoresis.
3) Succinate driven State 3 respiration was significantly inhibited both in vivo and in isolated mitochondria from the cortex of 3-NP treated rats, suggesting complex-I dysfunction in addition to inhibition of complex-II and succinate dehydrogenase activity contributes to cortico-striatal lesions in this model
Screening models for Parkinson & multiple sclerosisHimanshu Yadav
1. The document discusses various in vivo and in vitro models used to study Parkinson's disease (PD). In vivo models include antagonism of tremorine/oxotremorine-induced parkinsonism in mice, reserpine antagonism in mice, the MPTP model in monkeys, and circling behavior in nigrostriatal lesioned rats. In vitro methods include experiments using rat striatal slices and measuring dopamine stimulated adenyl cyclase activity.
2. The experimental autoimmune encephalomyelitis (EAE) model is described as the most widely used animal model for multiple sclerosis (MS). EAE is induced by immunizing mice with myelin antigens, causing an inflammatory de
By: Edward M. Barbieri and James Shorter
TDP-43 forms cytoplasmic aggregates in degenerating neurons of frontotemporal dementia (FTD) patients.
Laferrière et al. now establish that TDP-43 assemblies from distinct FTD subtypes have different structures,
neurotoxicities, and seeding activities, which correlate with FTD severity. Thus, distinct pathological TDP-43
assemblies akin to prion strains might underpin distinct FTD subtypes.
This study investigated the role of the complement receptor C5ar1 in epilepsy models. The researchers found that C5ar1 was upregulated in mouse models of status epilepticus. Treatment with the C5ar1 antagonist PMX53 reduced seizures in acute and chronic seizure models. PMX53's anticonvulsant effects were mediated by C5ar1, as they were not seen in C5ar1-deficient mice or with an inactive analogue. Inhibition of C5ar1 during status epilepticus lessened seizures, protected neurons from degeneration, and reduced mortality. The study suggests blocking C5ar1 activation may be a novel target for developing new anti-epileptic drugs.
1) Human growth hormone (HGH) treatment increased survival motor neuron (SMN) protein levels in human neuronal cells in a dose-dependent manner by activating the STAT5 signaling pathway.
2) Systemic administration of HGH to spinal muscular atrophy (SMA) mouse models induced higher levels of SMN protein in the brain and spinal cord.
3) HGH treatment improved disease phenotypes and increased survival in two severe SMA mouse models, confirming that activating the STAT5 pathway may be a promising therapeutic strategy for SMA.
The document discusses various strategies for overcoming resistance to endocrine therapy in advanced breast cancer, including targeting downstream signaling pathways like PI3K/Akt/mTOR that are often aberrantly activated in endocrine resistant disease. It reviews clinical trials demonstrating improved progression-free survival when adding drugs like everolimus or CDK4/6 inhibitors to hormonal therapy. The goal is to inhibit compensatory survival pathways that become relied upon by cancer cells to drive continued growth in the face of endocrine therapy.
This study found that both corticotropin (ACTH) and melanotropin (MSH) suppress the activation of human granulocytes and invertebrate immunocytes. ACTH required 2 hours to significantly inactivate human granulocytes, while MSH only required 20 minutes and was more potent. The addition of an enzyme inhibitor blocked the inactivation by ACTH, and tests found increased levels of MSH in samples incubated with ACTH, indicating ACTH is converted to MSH via an enzyme on granulocytes. Similar results were found with immune cells from mussels. The finding that HIV induces ACTH and MSH production suggests a role for these neuropeptides
Modulation Of Experimental Autoimmune Encephalomyelitis ByHoose71
VLA-2 antibody treatment significantly decreased clinical signs and pathology in a mouse model of multiple sclerosis called experimental autoimmune encephalomyelitis (EAE). Mice treated with a VLA-2 antibody during the effector phase, when symptoms first appeared, had lower disease severity scores and less demyelination in the spinal cord compared to control mice. VLA-2 antibody treatment was most effective when started immediately after disease onset, whereas treatment during induction or relapse phases did not significantly reduce EAE. The antibody may suppress EAE by blocking interactions between immune cells and the extracellular matrix that are important for inflammation in the central nervous system.
Temporal-Spatial Expressions of Spy1 in Rat Sciatic Nerve After CrushJiao Yang
1. The study examined the expression of the cell cycle protein Spy1 in a rat sciatic nerve crush injury model over time.
2. Spy1 expression was found to gradually increase after injury, peaking at day 3, due to increased expression in both axons and Schwann cells.
3. Spy1 expression correlated with Schwann cell proliferation after injury and Spy1 was found to localize in axons in the injured segment but did not co-localize with the growth protein GAP43.
The document discusses methylation processes and their role in human pathogenesis. It first describes how the MeCP2 protein functions in gene silencing through methyl-DNA binding and transcription repression, but that its role is more complex as a regulator rather than strict silencer of transcription. It also discusses how MeCP2 mutations cause Rett syndrome and interactions with other proteins. The role of DNA methylation in cancer is then covered, including global hypomethylation in cancer and hypermethylation of tumor suppressor genes. Finally, the potential of epigenetic therapies targeting DNA methyltransferases and histone deacetylases for cancer treatment is presented.
1) EGFR mutations are found in 10-30% of NSCLC cases and are associated with response to EGFR TKIs as first-line treatment.
2) Several studies including IPASS, EURTAC and LUX-Lung 3 have shown superior PFS for first-generation EGFR TKIs like gefitinib and erlotinib and the second-generation TKI afatinib compared to platinum-based chemotherapy as first-line treatment for EGFR mutation-positive NSCLC.
3) Meta-analyses of these studies demonstrate improved PFS for EGFR TKIs compared to chemotherapy in EGFR mutation-positive NSCLC, though OS is generally similar between the treatments
This study investigated the effects of pro-inflammatory cytokines TNFα and IFNγ and the hormone testosterone on expression of the chemokine receptors CX3CR1 and CCR2 on THP-1 monocytes. Flow cytometry analysis confirmed that TNFα and IFNγ increase cell surface expression of CX3CR1 and CCR2, while testosterone decreases expression of these receptors. Additionally, pre-treating cells with testosterone before cytokine exposure continued to decrease CX3CR1 and CCR2 expression levels compared to cytokine treatment alone. Future work will utilize controls to ensure the testosterone effects are receptor-mediated and not due to conversion to estrogen.
This study investigated the effects of lithium on the MEK-ERK signaling pathway in astrocytes and neurons. The key findings were:
1) In astrocytes, lithium decreased MEK and ERK phosphorylation in a time- and dose-dependent manner, inhibiting DNA synthesis and inducing cell cycle arrest.
2) In neurons, lithium enhanced MEK and ERK phosphorylation in a concentration-dependent way.
3) The opposing effects of lithium on the MEK-ERK pathway in astrocytes versus neurons suggest lithium may promote neural repair by inhibiting reactive gliosis while stimulating neurons.
Toll-like receptors (TLRs) are immune sensors that detect pathogen-associated molecular patterns and induce inflammatory responses. TLRs activate signaling cascades that lead to the production of cytokines and immune cell activation through adaptor proteins and kinases. Dysregulation of TLR signaling can cause or contribute to immune deficiencies, autoimmune diseases, and cancers. The document discusses TLR structure and function, downstream signaling pathways, regulation of the response, and diseases associated with TLR signaling abnormalities.
The document describes a study investigating whether ATM inhibitors can enhance the effectiveness of temozolomide (TMZ) in treating glioblastoma (GBM). The study found that the ATM inhibitor KU-55933 increased the sensitivity of inherently TMZ-sensitive GBM cell lines to TMZ, as shown by increased cell killing and G2/M cell cycle arrest when combined with TMZ. However, KU-55933 did not sensitize TMZ-resistant GBM cell lines to TMZ and did not induce G2/M arrest when combined with TMZ in these lines. These results suggest that ATM inhibition can improve the therapeutic effects of TMZ, but only in GBM cells that
This document summarizes information about the EGFR gene and the L858R mutation. It discusses how the EGFR gene codes for the epidermal growth factor receptor protein, which plays an important role in cancer signaling pathways. It specifically describes the L858R point mutation in the EGFR gene, which is common in non-small cell lung cancer. This mutation increases the tumor's sensitivity to EGFR inhibitors like gefitinib. Gefitinib is an ATP-competitive inhibitor that binds to the mutated EGFR protein and inhibits its kinase activity, providing a palliative treatment option for cancers with this mutation. A test was later developed to identify patients that may respond to gefitinib based on their tumor's
The document discusses various topics related to DNA methylation and its role in human pathology. It begins by describing the MeCP2 protein and its involvement in Rett syndrome through mutations in the MECP2 gene. It then discusses how alterations in DNA methylation levels and patterns are common in cancer and can lead to genomic instability. Finally, it examines the use of epigenetic therapies like DNA methyltransferase inhibitors and histone deacetylase inhibitors to treat cancer by reversing aberrant DNA methylation and histone modifications.
The document discusses various topics related to DNA methylation and its role in human pathology. It begins by describing the MeCP2 protein and its involvement in Rett syndrome through mutations in the MECP2 gene. It then discusses how alterations in DNA methylation levels and patterns are common in cancer and can lead to genomic instability. Finally, it examines the use of epigenetic therapies like DNA methyltransferase inhibitors and histone deacetylase inhibitors to treat cancer by reversing aberrant DNA methylation and histone modifications.
Antioxidant-mediated up-regulation of OGG1 via NRF2 induction is associated ...Enrique Moreno Gonzalez
Estrogen metabolism-mediated oxidative stress is suggested to play an important role in estrogen-induced breast carcinogenesis. We have earlier demonstrated that antioxidants,
vitamin C (Vit C) and butylated hydroxyanisole (BHA) inhibit 17β-estradiol (E2)-mediated oxidative stress and oxidative DNA damage, and breast carcinogenesis in female August
Copenhagen Irish (ACI) rats. The objective of the present study was to characterize the mechanism by which above antioxidants prevent DNA damage during breast carcinogenesis.
Decitabine Self Monitoring in Unstable Methylation of DNMT Patients: A Quasi ...Agriculture Journal IJOEAR
Abstract—
Background: Grey zone or intermediate zone CGG repeat in Pre-mutation FMR1 gene become in high prevalence in tropical rainforest area.
Problem: Bipolar disorder and Major depressive disorder used epigenetic drugs inhibitor to ameliorating their mood as well anticancer agent, decitabine are broadly used. Meanwhile, basic knowledge remains largely unknown.
Objective: Demethylation effect in grey zone methylation instability has to be controlled whereas up till now are to be disturbing the social behavior activities.
Hypothesis: Demethylation drive through, from >34 to <26 CGG repeat has behavior abnormalities.
Method: Quasi-Systematic Review with Bayesian network analysis using Science Direct and Ebsco-host search engine.
Result: One PRISMA Systematic Review flowchart to got the references and one table of 16 references to answer the methylation and demethylation in global living related to decitabine are recorded.
Discussion: Decitabine effect in epigenetic memory in mammals and neuro developmental, cognitive, behavioral and physical changes in grey zone and carrier permutation FMR1 gene are scanned.
Conclusion: Demethylation to high as well low grey zone CGG count could be self monitor due to instable methylation.
The political situation in Cambodia remains unstable as Prime Minister Hun Sen consolidates power by pressuring opposition parties and cracking down on dissent. Hun Sen's ruling Cambodian People's Party claims victory in the national election, but the opposition CNRP disputes the results. International observers note irregularities in the election process and an unlevel playing field due to the CPP's dominance over local media and resources.
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive function. Exercise causes chemical changes in the brain that may help protect against mental illness and improve symptoms for those who already suffer from conditions like depression and anxiety.
The document outlines a plan for shooting a film with the goal of hooking viewers and leaving them thinking. It recommends using attractive visuals and slowly revealing the truth of scenes to surprise viewers. It also suggests adding enigmas to make viewers think and adding mystery, as well as shocking viewers with a second body to confuse them. The final scene should incite fear and leave viewers thinking about the opening, indicating the film will be chilling.
This study found that both corticotropin (ACTH) and melanotropin (MSH) suppress the activation of human granulocytes and invertebrate immunocytes. ACTH required 2 hours to significantly inactivate human granulocytes, while MSH only required 20 minutes and was more potent. The addition of an enzyme inhibitor blocked the inactivation by ACTH, and tests found increased levels of MSH in samples incubated with ACTH, indicating ACTH is converted to MSH via an enzyme on granulocytes. Similar results were found with immune cells from mussels. The finding that HIV induces ACTH and MSH production suggests a role for these neuropeptides
Modulation Of Experimental Autoimmune Encephalomyelitis ByHoose71
VLA-2 antibody treatment significantly decreased clinical signs and pathology in a mouse model of multiple sclerosis called experimental autoimmune encephalomyelitis (EAE). Mice treated with a VLA-2 antibody during the effector phase, when symptoms first appeared, had lower disease severity scores and less demyelination in the spinal cord compared to control mice. VLA-2 antibody treatment was most effective when started immediately after disease onset, whereas treatment during induction or relapse phases did not significantly reduce EAE. The antibody may suppress EAE by blocking interactions between immune cells and the extracellular matrix that are important for inflammation in the central nervous system.
Temporal-Spatial Expressions of Spy1 in Rat Sciatic Nerve After CrushJiao Yang
1. The study examined the expression of the cell cycle protein Spy1 in a rat sciatic nerve crush injury model over time.
2. Spy1 expression was found to gradually increase after injury, peaking at day 3, due to increased expression in both axons and Schwann cells.
3. Spy1 expression correlated with Schwann cell proliferation after injury and Spy1 was found to localize in axons in the injured segment but did not co-localize with the growth protein GAP43.
The document discusses methylation processes and their role in human pathogenesis. It first describes how the MeCP2 protein functions in gene silencing through methyl-DNA binding and transcription repression, but that its role is more complex as a regulator rather than strict silencer of transcription. It also discusses how MeCP2 mutations cause Rett syndrome and interactions with other proteins. The role of DNA methylation in cancer is then covered, including global hypomethylation in cancer and hypermethylation of tumor suppressor genes. Finally, the potential of epigenetic therapies targeting DNA methyltransferases and histone deacetylases for cancer treatment is presented.
1) EGFR mutations are found in 10-30% of NSCLC cases and are associated with response to EGFR TKIs as first-line treatment.
2) Several studies including IPASS, EURTAC and LUX-Lung 3 have shown superior PFS for first-generation EGFR TKIs like gefitinib and erlotinib and the second-generation TKI afatinib compared to platinum-based chemotherapy as first-line treatment for EGFR mutation-positive NSCLC.
3) Meta-analyses of these studies demonstrate improved PFS for EGFR TKIs compared to chemotherapy in EGFR mutation-positive NSCLC, though OS is generally similar between the treatments
This study investigated the effects of pro-inflammatory cytokines TNFα and IFNγ and the hormone testosterone on expression of the chemokine receptors CX3CR1 and CCR2 on THP-1 monocytes. Flow cytometry analysis confirmed that TNFα and IFNγ increase cell surface expression of CX3CR1 and CCR2, while testosterone decreases expression of these receptors. Additionally, pre-treating cells with testosterone before cytokine exposure continued to decrease CX3CR1 and CCR2 expression levels compared to cytokine treatment alone. Future work will utilize controls to ensure the testosterone effects are receptor-mediated and not due to conversion to estrogen.
This study investigated the effects of lithium on the MEK-ERK signaling pathway in astrocytes and neurons. The key findings were:
1) In astrocytes, lithium decreased MEK and ERK phosphorylation in a time- and dose-dependent manner, inhibiting DNA synthesis and inducing cell cycle arrest.
2) In neurons, lithium enhanced MEK and ERK phosphorylation in a concentration-dependent way.
3) The opposing effects of lithium on the MEK-ERK pathway in astrocytes versus neurons suggest lithium may promote neural repair by inhibiting reactive gliosis while stimulating neurons.
Toll-like receptors (TLRs) are immune sensors that detect pathogen-associated molecular patterns and induce inflammatory responses. TLRs activate signaling cascades that lead to the production of cytokines and immune cell activation through adaptor proteins and kinases. Dysregulation of TLR signaling can cause or contribute to immune deficiencies, autoimmune diseases, and cancers. The document discusses TLR structure and function, downstream signaling pathways, regulation of the response, and diseases associated with TLR signaling abnormalities.
The document describes a study investigating whether ATM inhibitors can enhance the effectiveness of temozolomide (TMZ) in treating glioblastoma (GBM). The study found that the ATM inhibitor KU-55933 increased the sensitivity of inherently TMZ-sensitive GBM cell lines to TMZ, as shown by increased cell killing and G2/M cell cycle arrest when combined with TMZ. However, KU-55933 did not sensitize TMZ-resistant GBM cell lines to TMZ and did not induce G2/M arrest when combined with TMZ in these lines. These results suggest that ATM inhibition can improve the therapeutic effects of TMZ, but only in GBM cells that
This document summarizes information about the EGFR gene and the L858R mutation. It discusses how the EGFR gene codes for the epidermal growth factor receptor protein, which plays an important role in cancer signaling pathways. It specifically describes the L858R point mutation in the EGFR gene, which is common in non-small cell lung cancer. This mutation increases the tumor's sensitivity to EGFR inhibitors like gefitinib. Gefitinib is an ATP-competitive inhibitor that binds to the mutated EGFR protein and inhibits its kinase activity, providing a palliative treatment option for cancers with this mutation. A test was later developed to identify patients that may respond to gefitinib based on their tumor's
The document discusses various topics related to DNA methylation and its role in human pathology. It begins by describing the MeCP2 protein and its involvement in Rett syndrome through mutations in the MECP2 gene. It then discusses how alterations in DNA methylation levels and patterns are common in cancer and can lead to genomic instability. Finally, it examines the use of epigenetic therapies like DNA methyltransferase inhibitors and histone deacetylase inhibitors to treat cancer by reversing aberrant DNA methylation and histone modifications.
The document discusses various topics related to DNA methylation and its role in human pathology. It begins by describing the MeCP2 protein and its involvement in Rett syndrome through mutations in the MECP2 gene. It then discusses how alterations in DNA methylation levels and patterns are common in cancer and can lead to genomic instability. Finally, it examines the use of epigenetic therapies like DNA methyltransferase inhibitors and histone deacetylase inhibitors to treat cancer by reversing aberrant DNA methylation and histone modifications.
Antioxidant-mediated up-regulation of OGG1 via NRF2 induction is associated ...Enrique Moreno Gonzalez
Estrogen metabolism-mediated oxidative stress is suggested to play an important role in estrogen-induced breast carcinogenesis. We have earlier demonstrated that antioxidants,
vitamin C (Vit C) and butylated hydroxyanisole (BHA) inhibit 17β-estradiol (E2)-mediated oxidative stress and oxidative DNA damage, and breast carcinogenesis in female August
Copenhagen Irish (ACI) rats. The objective of the present study was to characterize the mechanism by which above antioxidants prevent DNA damage during breast carcinogenesis.
Decitabine Self Monitoring in Unstable Methylation of DNMT Patients: A Quasi ...Agriculture Journal IJOEAR
Abstract—
Background: Grey zone or intermediate zone CGG repeat in Pre-mutation FMR1 gene become in high prevalence in tropical rainforest area.
Problem: Bipolar disorder and Major depressive disorder used epigenetic drugs inhibitor to ameliorating their mood as well anticancer agent, decitabine are broadly used. Meanwhile, basic knowledge remains largely unknown.
Objective: Demethylation effect in grey zone methylation instability has to be controlled whereas up till now are to be disturbing the social behavior activities.
Hypothesis: Demethylation drive through, from >34 to <26 CGG repeat has behavior abnormalities.
Method: Quasi-Systematic Review with Bayesian network analysis using Science Direct and Ebsco-host search engine.
Result: One PRISMA Systematic Review flowchart to got the references and one table of 16 references to answer the methylation and demethylation in global living related to decitabine are recorded.
Discussion: Decitabine effect in epigenetic memory in mammals and neuro developmental, cognitive, behavioral and physical changes in grey zone and carrier permutation FMR1 gene are scanned.
Conclusion: Demethylation to high as well low grey zone CGG count could be self monitor due to instable methylation.
The political situation in Cambodia remains unstable as Prime Minister Hun Sen consolidates power by pressuring opposition parties and cracking down on dissent. Hun Sen's ruling Cambodian People's Party claims victory in the national election, but the opposition CNRP disputes the results. International observers note irregularities in the election process and an unlevel playing field due to the CPP's dominance over local media and resources.
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive function. Exercise causes chemical changes in the brain that may help protect against mental illness and improve symptoms for those who already suffer from conditions like depression and anxiety.
The document outlines a plan for shooting a film with the goal of hooking viewers and leaving them thinking. It recommends using attractive visuals and slowly revealing the truth of scenes to surprise viewers. It also suggests adding enigmas to make viewers think and adding mystery, as well as shocking viewers with a second body to confuse them. The final scene should incite fear and leave viewers thinking about the opening, indicating the film will be chilling.
The document outlines a student film project about a boy who experiences the transition from childhood to adulthood. The story follows the boy who lives happily at home until his 18th birthday, when his mother gives him a light switch that allows him to see the outside world in black and white. When he switches it on and leaves home, he is initially fascinated by his new independence but soon finds that the realities of adulthood are bleak and unforgiving. The film aims to symbolize the tensions between childhood and adulthood using color schemes and the boy's journey outside of the home. Locations will include the student's house and surrounding streets, and he will play the main character with minimal costume needs.
The document discusses where real creativity can be found in media such as films and music. It argues that true creativity is often seen in independent and alternative works, rather than big budget blockbusters or popular mainstream works. As examples, it cites creative films like Boyhood and Eternal Sunshine of the Spotless Mind, as well as imaginative musicians like Flying Lotus and Grizzly Bear. While the digital age has opened up more opportunities for creativity, it can also be difficult to be truly original given how much media is produced. The document then analyzes five short films that demonstrate creativity through their stories, visuals, meanings, and editing techniques.
The political situation in Cambodia remains volatile as Prime Minister Hun Sen consolidates power by imprisoning opposition leaders and cracking down on dissent. Long-time opposition leader Kem Sokha was arrested in 2017 on treason charges widely seen as politically motivated. Without a clear challenger, Hun Sen is expected to extend his 33 year rule in the upcoming national elections later this year, though international observers warn the conditions may not allow for a free and fair vote.
The political situation in Cambodia remains unstable as Prime Minister Hun Sen consolidates power by pressuring opposition parties and cracking down on dissent. Hun Sen's ruling Cambodian People's Party claimed victory in the most recent national election, though the election was marred by accusations of voter intimidation and suppression of opposition voices. Critics fear that Cambodia is slipping deeper into authoritarianism under Hun Sen's increasingly authoritarian rule.
The document summarizes editing a photo in Photoshop. The photo is separated into two layers - the flower and background. The background layer has its saturation decreased to make it black and white while keeping the flower in color. A wind filter is added to the background layer to give a dreamlike, fading effect that ties into the film's aesthetic. Editing is then done on the flower layer to make it more vibrant before moving on to poster creation.
The document describes the steps taken to create a movie poster. First, the creator opened a new document and imported the edited photo, sizing it appropriately. They created a cream-colored background layer and faded out part of the photo using a layer mask to make space for additional elements. Next, the creator added the movie title in a bold black font with pink O's to match the flower, and a tagline "Growing up isn't all Black and White" with inverted colors. Finally, release date and credit text were added at the bottom in faded grey.
The political situation in Cambodia remains volatile as Prime Minister Hun Sen consolidates power by pressuring opposition parties and cracking down on dissent. Hun Sen's ruling Cambodian People's Party claims victory in the national election despite allegations of voter intimidation and fraud. Critics accuse Hun Sen of dismantling democracy in Cambodia and establishing a de facto one-party state.
JC Fall 2009-Role of TNFalpha-Converting Enzyme (TACE) Inhibition on Amyloid ...Nisha Rizvi
This document summarizes the findings of a study investigating the effects of BMS-561392, a TNFα converting enzyme (TACE) inhibitor, on amyloid beta (Aβ) production. The study found that BMS-561392 selectively inhibited TACE without affecting BACE or increasing Aβ levels. This suggests that while TACE and BACE reside in the same compartment, they do not compete for the same pool of amyloid precursor protein and target distinct pathways. Therefore, TACE-targeted anti-inflammatory drugs may not increase Aβ generation in Alzheimer's disease patients as previously thought. The authors conclude that BMS-561392 is an unlikely AD drug candidate but that further research into centrally-acting
Seminar-Spring 2010-Role of Wnt signaling in Alzheimer's disease pathogenesisNisha Rizvi
1. Wnt ligands are involved in pre- and postsynaptic protein clustering and assembly at synapses and form functional new synapses.
2. Activation of Wnt signaling protects against Aβ toxicity in Alzheimer's disease by increasing neuronal survival, decreasing apoptosis, and stabilizing β-catenin levels. Inhibition of Wnt signaling can cause neuronal damage.
3. Wnt signaling interacts and forms a crosstalk with other pathways important in Alzheimer's like M1 muscarinic acetylcholine receptor activation and PPARγ activation, which may provide neuroprotective effects against Aβ toxicity.
The document discusses a study investigating the effects of food restriction on gene expression. It was previously found that 15 genes were upregulated in the brain during food restriction, suggesting they are part of an ancient stress response pathway. The current study aims to test if these genes are also induced in other tissue types under food restriction. Mice were either food restricted or not for 5 days, then gene expression was analyzed using qPCR in various tissues including the kidney. It was found that Angptl4, Mertk, Arrdc2 and Cdkn1a were significantly upregulated in the kidney of food restricted mice compared to controls, providing further evidence they are part of a general stress response pathway activated by food restriction across multiple
1) The study demonstrates that phosphorylation of huntingtin protein at serine 421 (S421) protects against toxicity caused by polyglutamine expansion in huntingtin, the cause of Huntington's disease.
2) Calcineurin (CaN), a calcium-activated phosphatase, was found to dephosphorylate S421 both in vitro and in cells. Inhibition of CaN using FK506 increased S421 phosphorylation and prevented polyQ-huntingtin toxicity in neuronal cells.
3) Administration of FK506 to mice also increased phosphorylation of endogenous huntingtin at S421 in the brain. Therefore, inhibition of CaN's dephosphorylation of S421 may have therapeutic potential for treating Huntington's
PTC124 is a compound that was discovered to suppress premature stop codons caused by nonsense mutations in mRNA. It was found to induce read-through of premature stop codons in in vitro and cell culture experiments involving luciferase reporter genes containing premature stop codons. In mouse and human studies, PTC124 treatment led to increased production of full-length dystrophin protein in Duchenne muscular dystrophy patients and mdx mice, demonstrating its ability to induce read-through of the premature stop codon in the dystrophin gene. The mechanism of action is unknown but PTC124 appears to induce read-through rather than protein stabilization.
AZD3293 is a novel BACE1 inhibitor being developed for the treatment of Alzheimer's disease. This document reports results from two Phase I clinical trials of AZD3293. The first was a single ascending dose study that evaluated doses from 1-750 mg in healthy young and elderly subjects and assessed food effects. The second was a multiple ascending dose study that administered AZD3293 at doses of 15-150 mg once daily or 70 mg once weekly for 2 weeks to elderly healthy subjects and patients with mild-moderate Alzheimer's disease. AZD3293 was generally well tolerated. Pharmacokinetic results showed dose-dependent increases in drug levels and half-lives of 16-21 hours. AZD3293 significantly reduced
This document summarizes research on Tuberous Sclerosis Complex (TSC), including key facts about the disease, progress that has been made in understanding the genetics and molecular mechanisms, development of treatments like Everolimus, and ongoing areas of research focus like clinical trials of new drugs and studying disease mechanisms using cellular and animal models.
This document contains summaries of 4 poster presentations from a conference on Alzheimer's disease and dementia research.
The first poster discusses findings that the ER stress transcription factor XBP1s protects against amyloid-beta neurotoxicity in a fruit fly model of Alzheimer's and in human neuroblastoma cells. XBP1s prevents the accumulation of free calcium in the cytosol, explaining its neuroprotective effects.
The second poster reports that amyloid-beta oligomers impair neuronal function and morphology in rat and mouse neuronal cultures and hippocampal slices through a mechanism linked to NMDA receptor signaling and activation of the protein Jacob.
The third poster examines the role of O-GlcNAc glycosylation in hippocampal synaptic
Publication du Pr. Baulieu dans PNAS - Avril 2014InstitutBaulieu
FKBP52 interacts directly with the pathological Tau-P301L mutant and induces its oligomerization and assembly into filaments in vitro. In a zebrafish model of tauopathy expressing human Tau-P301L, knockdown of FKBP52 is sufficient to restore defective axonal outgrowth and branching in spinal motoneurons caused by Tau-P301L expression. FKBP52 appears to modulate the pathological activity of Tau-P301L both in vitro and in vivo.
This document announces a journal club presentation on the effects of inhibiting TNFα-converting enzyme (TACE) on amyloid β production and amyloid precursor protein (APP) processing in vitro and in vivo. TACE inhibition has potential to alleviate TNFα-mediated inflammation in Alzheimer's disease by decreasing soluble TNFα levels. However, TACE also cleaves APP and its inhibition could increase amyloidogenic APP processing. The presented study demonstrates that selective TACE inhibition using BMS-561392 reduces APPα levels without increasing amyloid β production, suggesting TACE inhibition may benefit Alzheimer's disease by reducing neuroinflammation without exacerbating amyloid pathology.
Normal human prostate epithelial cells (PrEC) are sensitive to TRAIL-induced apoptosis, contrary to previous beliefs. TRAIL treatment of PrEC leads to DNA fragmentation and cell death. While PrEC are sensitive, other primary cell types like fibroblasts and endothelial cells are resistant. PrEC contain fewer decoy receptors that inhibit TRAIL's apoptotic signal compared to resistant cells. Inhibition of protein synthesis enhances TRAIL's toxicity in PrEC, suggesting short-lived proteins inhibit aspects of TRAIL-induced apoptosis in these cells. This has implications for using TRAIL as a potential prostate cancer treatment.
This document describes an experiment examining the effect of the PI3K/AKT pathway on HOXA1 expression in prostate cancer stem-like cells induced by diepoxybutane (DEB). DU145 prostate cancer cells were treated with DEB to generate cancer stem cells. RNA and protein were extracted from treated and untreated cells. PCR and gel electrophoresis showed expression of HOXA1, p38, cMYC, and JNK-2 genes in DEB-treated cells. Western blot and immunofluorescence were used to analyze protein expression and localization of HOXA1, p53, and pro-caspase 9. Results indicate DEB activates the PI3K/AKT pathway, leading to changes in
1. Researchers identified a novel isoform of the tumor suppressor gene DLC1 called DLC1-i4 through 5' RACE. DLC1-i4 encodes a 1125 amino acid protein with a distinct N-terminus compared to other DLC1 isoforms.
2. DLC1-i4 expression is frequently downregulated in multiple carcinoma cell lines, including esophageal, gastric, breast, colorectal, cervical and lung cancers. Its promoter region contains CpG islands that are often methylated, silencing expression.
3. Ectopic expression of DLC1-i4 in silenced tumor cells strongly inhibited their growth and colony formation, demonstrating its tumor suppressive
This study examines the correlation between cytoplasmic p21 levels, AKT activation, and response to tamoxifen in breast cancer patients. The results show that cytoplasmic p21 levels strongly correlate with pAKT levels and predict a poor response to tamoxifen therapy. In vitro experiments demonstrate that the growth factor HRG β 1 induces both nuclear and cytoplasmic p21 expression in breast cancer cells, impairing the inhibitory effects of tamoxifen on cell cycle progression and apoptosis.
This document is a thesis examining the effects of recombinant adeno-associated viral arginase 1 in transgenic mice with tau pathology. The introduction discusses Alzheimer's disease and tauopathies, L-arginine metabolism and associated pathways, and the mouse model used. The goal was to identify the effects of overexpressing arginase 1 in hippocampal neurons using viral vectors in this mouse model. Histological experiments included staining for various proteins and markers to analyze the effects.
TP receptors augment cellular immune responses and inflammatory tissue injury. Mice lacking the TP receptor, which binds thromboxane A2, show reduced T cell proliferation in response to mitogens and alloantigens. They also show attenuated tissue injury in a cardiac transplant model of inflammation. Thus, thromboxane signaling through the TP receptor enhances immune responses and inflammation, suggesting that specific inhibition of this receptor may reduce inflammation without the side effects of broad COX inhibition by NSAIDs.
Tetrahydrohyperforin (IDN5706), a derivative of the active molecule hyperforin in St. John's Wort, was examined for its ability to prevent cognitive deficits and synaptic impairment in an Alzheimer's disease mouse model. Five-month old APPswe/PSEN1DE9 mice were treated with IDN5706 for 10 weeks. IDN5706 improved memory, prevented decreases in synaptic proteins and LTP, and reduced amyloid-beta plaque burden, tau hyperphosphorylation, and astrogliosis. In cell cultures, IDN5706 decreased amyloid precursor protein processing leading to amyloid-beta peptide generation. The results suggest IDN5706 may be a potential therapeutic for treating Alzheimer's
Tetrahydrohyperforin (IDN5706), a derivative of the active molecule hyperforin in St. John's Wort, was examined for its ability to prevent cognitive deficits and synaptic impairment in an Alzheimer's disease mouse model. Five-month old APPswe/PSEN1DE9 mice were treated with IDN5706 for 10 weeks. IDN5706 improved memory, prevented decreases in synaptic proteins and LTP, and reduced amyloid-beta plaque burden, tau hyperphosphorylation, and astrogliosis. In cell cultures, IDN5706 decreased amyloid precursor protein processing leading to amyloid-beta peptide generation. The results suggest IDN5706 may be a potential therapeutic for treating Alzheimer's
Endothelial Cell Mediated Delay of Blood Brain Barrier Recovery Following Tra...Arthur Stem
TBI is the leading cause of death among young adults and children in the developed world, accounting for over 50,000 deaths per year. [12] TBI results in a sleuth of poor health outcomes, including hemorrhaging, seizures, neural edema, neural inflammation, and cognitive and emotional disabilities. All of these outcomes are a direct result of fundamental degradation of the BBB over a time course post TBI. [1] [12] The BBB is an integral structure that forms around the microvascular of the cerebral cavity. Endothelial cells form the basal membrane through which strictly controlled movement of molecules is observed between the extravascular and intravascular space across this basal membrane. This basal membrane is maintained by endothelial cells, having tight junctions between them to make up the pores through which transport of molecules can occur between the brain and microvasculature. These tight junctions are maintained through cross-talk between the endothelial cells and supporting neurons such as astrocytes and pericytes. [2] A multitude of proteins make up the tight junctions between the endothelial cells, including six main scaffolding structures Claudins 1, 3, and 5, ZO-1, Occludins, and Cadherins. [3] VEGF release following trauma induces endothelial cells to release matrix metalloproteinases (MMPs), in particular MMP9, which can catalyze the N-terminal amino acids that compose the tight junction protein ZO-1. [10] [11] MMP9 when in circulation is also known to activate tumor necrosis factor alpha (TNFɑ) which in turn upregulates transcription of MMP9, creating a positive feedback loop. [11] The management of MMP production is three fold, transcription, proenzyme activation, and substrate inhibition. [11] In our study, it is proenzyme activation via TP that is the focus and how that affects the overall transcription levels of the tight junction proteins within the endothelial cells and astrocytes.
This document summarizes a book titled "Role of Plants, environmental toxins and physical neurotoxicological factors in Amyotrophic lateral sclerosis, Alzheimer Disease and Other Neurodegenerative Diseases." It has 10 authors from various countries. The book aims to observe the effects of plant neurotoxins, physical factors, and geography on neurodegenerative diseases like ALS, PD, and AD. It reviews literature on geographic clusters of these diseases and genetic mutations. It also discusses the brain's high metabolic needs, interactions between neurons and astrocytes, and how impairments in these systems can lead to neurodegeneration. The book presents figures and intends to develop new hypotheses and therapies based on contributing pathogenic factors.
2011 - Cellular inhibitor of apoptosis protein-1 (cIAP1) can regulate E2F1 tr...Simon Gemble
Cellular inhibitor of apoptosis protein-1 (cIAP1) can directly interact with the transcription factor E2F1 and increase its transcriptional activity. cIAP1 is recruited to E2F1 binding sites on cyclin E and cyclin A promoters in a cell cycle-dependent manner. Silencing cIAP1 inhibits E2F1 DNA binding and transcriptional activation of cyclin E, reducing cell proliferation. Thus, one function of nuclear cIAP1 is to regulate E2F1 transcriptional activity and control cell cycle progression.
1) The researchers generated mutations of the I-F11 nerve agent bioscavenger enzyme that eliminated glycosylation by mutating asparagine residues 253 and 324.
2) Testing showed the double mutant had increased thermal stability in vitro but no change in circulatory half-life in vivo compared to the glycosylated wild type.
3) Mutation of either residue 253 or 324 alone resulted in partial deglycosylation, while mutation of both residues eliminated glycosylation, indicating these are the sites of glycosylation in the enzyme.
हिंदी वर्णमाला पीपीटी, hindi alphabet PPT presentation, hindi varnamala PPT, Hindi Varnamala pdf, हिंदी स्वर, हिंदी व्यंजन, sikhiye hindi varnmala, dr. mulla adam ali, hindi language and literature, hindi alphabet with drawing, hindi alphabet pdf, hindi varnamala for childrens, hindi language, hindi varnamala practice for kids, https://www.drmullaadamali.com
Strategies for Effective Upskilling is a presentation by Chinwendu Peace in a Your Skill Boost Masterclass organisation by the Excellence Foundation for South Sudan on 08th and 09th June 2024 from 1 PM to 3 PM on each day.
LAND USE LAND COVER AND NDVI OF MIRZAPUR DISTRICT, UPRAHUL
This Dissertation explores the particular circumstances of Mirzapur, a region located in the
core of India. Mirzapur, with its varied terrains and abundant biodiversity, offers an optimal
environment for investigating the changes in vegetation cover dynamics. Our study utilizes
advanced technologies such as GIS (Geographic Information Systems) and Remote sensing to
analyze the transformations that have taken place over the course of a decade.
The complex relationship between human activities and the environment has been the focus
of extensive research and worry. As the global community grapples with swift urbanization,
population expansion, and economic progress, the effects on natural ecosystems are becoming
more evident. A crucial element of this impact is the alteration of vegetation cover, which plays a
significant role in maintaining the ecological equilibrium of our planet.Land serves as the foundation for all human activities and provides the necessary materials for
these activities. As the most crucial natural resource, its utilization by humans results in different
'Land uses,' which are determined by both human activities and the physical characteristics of the
land.
The utilization of land is impacted by human needs and environmental factors. In countries
like India, rapid population growth and the emphasis on extensive resource exploitation can lead
to significant land degradation, adversely affecting the region's land cover.
Therefore, human intervention has significantly influenced land use patterns over many
centuries, evolving its structure over time and space. In the present era, these changes have
accelerated due to factors such as agriculture and urbanization. Information regarding land use and
cover is essential for various planning and management tasks related to the Earth's surface,
providing crucial environmental data for scientific, resource management, policy purposes, and
diverse human activities.
Accurate understanding of land use and cover is imperative for the development planning
of any area. Consequently, a wide range of professionals, including earth system scientists, land
and water managers, and urban planners, are interested in obtaining data on land use and cover
changes, conversion trends, and other related patterns. The spatial dimensions of land use and
cover support policymakers and scientists in making well-informed decisions, as alterations in
these patterns indicate shifts in economic and social conditions. Monitoring such changes with the
help of Advanced technologies like Remote Sensing and Geographic Information Systems is
crucial for coordinated efforts across different administrative levels. Advanced technologies like
Remote Sensing and Geographic Information Systems
9
Changes in vegetation cover refer to variations in the distribution, composition, and overall
structure of plant communities across different temporal and spatial scales. These changes can
occur natural.
This presentation includes basic of PCOS their pathology and treatment and also Ayurveda correlation of PCOS and Ayurvedic line of treatment mentioned in classics.
How to Manage Your Lost Opportunities in Odoo 17 CRMCeline George
Odoo 17 CRM allows us to track why we lose sales opportunities with "Lost Reasons." This helps analyze our sales process and identify areas for improvement. Here's how to configure lost reasons in Odoo 17 CRM
The simplified electron and muon model, Oscillating Spacetime: The Foundation...RitikBhardwaj56
Discover the Simplified Electron and Muon Model: A New Wave-Based Approach to Understanding Particles delves into a groundbreaking theory that presents electrons and muons as rotating soliton waves within oscillating spacetime. Geared towards students, researchers, and science buffs, this book breaks down complex ideas into simple explanations. It covers topics such as electron waves, temporal dynamics, and the implications of this model on particle physics. With clear illustrations and easy-to-follow explanations, readers will gain a new outlook on the universe's fundamental nature.
How to Setup Warehouse & Location in Odoo 17 InventoryCeline George
In this slide, we'll explore how to set up warehouses and locations in Odoo 17 Inventory. This will help us manage our stock effectively, track inventory levels, and streamline warehouse operations.
বাংলাদেশের অর্থনৈতিক সমীক্ষা ২০২৪ [Bangladesh Economic Review 2024 Bangla.pdf] কম্পিউটার , ট্যাব ও স্মার্ট ফোন ভার্সন সহ সম্পূর্ণ বাংলা ই-বুক বা pdf বই " সুচিপত্র ...বুকমার্ক মেনু 🔖 ও হাইপার লিংক মেনু 📝👆 যুক্ত ..
আমাদের সবার জন্য খুব খুব গুরুত্বপূর্ণ একটি বই ..বিসিএস, ব্যাংক, ইউনিভার্সিটি ভর্তি ও যে কোন প্রতিযোগিতা মূলক পরীক্ষার জন্য এর খুব ইম্পরট্যান্ট একটি বিষয় ...তাছাড়া বাংলাদেশের সাম্প্রতিক যে কোন ডাটা বা তথ্য এই বইতে পাবেন ...
তাই একজন নাগরিক হিসাবে এই তথ্য গুলো আপনার জানা প্রয়োজন ...।
বিসিএস ও ব্যাংক এর লিখিত পরীক্ষা ...+এছাড়া মাধ্যমিক ও উচ্চমাধ্যমিকের স্টুডেন্টদের জন্য অনেক কাজে আসবে ...
it describes the bony anatomy including the femoral head , acetabulum, labrum . also discusses the capsule , ligaments . muscle that act on the hip joint and the range of motion are outlined. factors affecting hip joint stability and weight transmission through the joint are summarized.
Executive Directors Chat Leveraging AI for Diversity, Equity, and InclusionTechSoup
Let’s explore the intersection of technology and equity in the final session of our DEI series. Discover how AI tools, like ChatGPT, can be used to support and enhance your nonprofit's DEI initiatives. Participants will gain insights into practical AI applications and get tips for leveraging technology to advance their DEI goals.
2. Kim et al. • TACE Inhibition Does Not Enhance A Production
Figure 1. BMS-561392 inhibits secretion of sAPP␣ and TNF␣ in a dose-dependent manner.
CHO cells expressing human APP695wt and pro-TNF␣ were treated with BMS-561392 and
TAPI-I for 24 h. A, Inhibition of TNF␣ secretion by TAPI-I and BMS-561392. Inhibition of 50%
with respect to DMSO controls was achieved with 0.90 M TAPI-I and 0.15 M BMS-561392. B,
Inhibition of sAPP␣ secretion by TAPI-I and BMS-561392 from CHO cells expressing APPwt and
APPswe. Inhibition of 50% with respect to DMSO controls was achieved with 59.1 M TAPI-I and
4.47 M BMS-561392 on CHO-APPwt cells, and 0.23 M BMS-561392 on CHO-APPswe cells.
Data are expressed as the mean of three independent experiments ϮSE measurements. *p Ͻ
0.01, two-tailed Student’s t test; **, ***p Ͻ 0.0001, two-tailed Student’s t test.
ADAM-10 and ADAM-9 (Lammich et al., 1999; Koike et al.,
1999), because of the lack of a specific inhibitor.
BMS-561392 is a novel, potent and highly selective TACE inhibitor (supplemental Fig. 1, Table 1, available at www.
jneurosci.org as supplemental material) that effectively suppresses excessive TNF␣ secretion. In this study, we have characterized the effects of BMS-561392 on TACE and BACE cleavage
of APP in vitro. We also infused the compound into the brain of
human APP-expressing Tg2576 and wild-type mice to examine
its effects on APP processing in vivo. Finally, we have addressed
the question of whether TACE and BACE compete for APP substrate. Our data support the notion that TACE and BACE share
the same pool of APP, but do not normally compete for APP
substrate.
Materials and Methods
Cell culture and drug treatment. Chinese hamster ovary (CHO) cells stably transfected with the wild-type 695- amino acid isoform of APP (APPwt) and a familial AD-associated APP Swedish mutant (APPswe) (Forman et al., 1997) as well as APP-trans-Golgi network (APP-TGN)
(Chyung et al., 1997; Skovronsky et al., 2000; Huse et al., 2002) were
grown in ␣-MEM (Invitrogen) containing 5% fetal bovine serum, penicillin/streptomycin, and 50 g/ml G418 according to standard protocols.
J. Neurosci., November 12, 2008 • 28(46):12052–12061 • 12053
Cortices from neonatal Tg2576 mouse brains were isolated and incubated in 0.1% trypsin/HBSS/0.5 mM EDTA without Ca 2ϩ or Mg 2ϩ (Invitrogen), and cells were mechanically dissociated using a fire-polished
pipette. Cells were plated in Dulbecco’s minimum essential medium
(DMEM) plus 10% FBS in poly-D-lysine-coated six-well plates at a density of 0.5 ϫ 10 6 cells/well. Three hours after plating, medium was replaced with DMEM plus B27 supplements (Invitrogen) to promote neuronal survival and inhibit growth of non-neuronal cells. Neurons were
used for experiments after 7 d in vitro. CHO-pro-TNF␣ cells were generated by transfection of pro-TNF␣ vector with alkaline phosphatase
tagged at its C-terminal (a gift of Dr. Carl Blobel, Hospital for Special
Surgery, New York, NY) (Chesneau et al., 2003) into CHO-Pro5, using
Nucleofector (Amaxa). After subcloning using Zeocin as a selection
agent, they were grown under the same conditions as other CHO cells. CHO
cells were plated in six-well plates at a density of 6.75 ϫ 10 5 cells/well for drug
treatment. 24 h after plating, medium was replaced with BMS-561392
(Bristol-Myers Squibb), TAPI-I [(N-R-(2-hydroxyaminocarbonyl)methyl]4-methylpentanoyl-L-naphthylanyl-L-alanine 2 aminoethyl amide-I; Peptide International), or the BACE inhibitor III (Merck; the molecular structure is proprietary information of Merck) in media containing 0.1% DMSO
with or without 10 M PMA (Phorbol 12-myristate 13-acetate; Sigma). Control wells received media with 0.1% DMSO only.
Osmotic pump implantation and drug treatment in vivo. The protocol
of mouse brain hippocampal microinfusion have been carefully reviewed
and approved by the Institutional Animal Care and Use Committee
(IACUC) at University of Pennsylvania. Briefly, Alzet pumps (type 2004,
100 l, 0.25 l/hr; Alzet Osmotic Pumps ) were filled with BMS-561392
at concentration of 8 mg/ml in the citric acid, pH 3.5. A broad-range
matrix metalloprotease (MMP) inhibitor TAPI-I was prepared at the
same molar concentration. The osmotic pumps then were kept in 0.9%
saline at 37°C before implantation. Three groups of six to eight month
old Tg2576 and wild-type mice (n ϭ 5 per group) were used in this study.
Mice were anesthetized by an intraperitoneal injection of ketamine hydrochloride (1 mg/10 g) and xylazine (0.1 mg/10 g), and canula was
stereotactically inserted into the right lateral ventricle using sterotaxic
coordinates (anterior–posterior, 0.5 mm; medial–lateral, 1.0 mm; dorsal–ventral, 2.7–3.0 mm; David Kopf Instrument). Instant dental cement
was used to secure canula to skull. Osmotic pump was placed subcutaneously on the back of mice. After surgery, mice were housed individually and infusion lasted for 14 d.
The mice behaved normally during treatment. Cerebrospinal fluid
(CSF) was collected while mice were under anesthesia, and 〈40/42
levels were measured by sandwich ELISA. Mice were subsequently killed
and perfused with PBS, and the cortex and hippocampus from the drug/
vehicle-infused right hemisphere were dissected, immediately frozen in
dry ice, and kept at Ϫ80°C for further processing.
Tg2576 mice were maintained on a C57B6/SJL F2 background by
successive backcrossing to wild-type C57B6/SJL F1 females. All mice
were generated and handled according to protocols approved by the
University of Pennsylvania Institutional Animal Care and Use Committee guidelines.
Sample processing and Western blot analysis. Cell lysates were collected
in RIPA buffer (0.5% sodium deoxycholate, 0.1% SDS, 1% Nonidet
P-40, 5 mM EDTA in TBS, pH 8.0) in the presence of protease inhibitors
(1 g/ml each of pepstatin A, leupeptin, L-1-tosylamido-2-phenylethyl
chloromethyl ketone, 1-chloro-3-tosylamido-7-amino-2-heptanone,
soybean trypsin inhibitor, and 0.5 mM phenylmethylsulfonyl fluoride)
and briefly sonicated. Protease inhibitors were also added to conditioned
media samples.
Frozen brains of Tg2576 mice were directly lysed in RIPA buffer (150
mg/ml) containing protease inhibitors, sonicated, and centrifuged at
100,000 ϫ g for 20 min at 4°C. The supernatants were saved and used for
A sandwich ELISA analysis and Western blotting. To selectively extract
A, sAPP␣, and sAPP, wild-type mouse brains were lysed in 0.2%
DEA-containing 50 mM NaCl, homogenized using a pestle, and centrifuged at 100,000 ϫ g for 1 h at 4°C (Miller et al., 2003). The supernatants
were saved and neutralized by adding 1/10th volume of 0.5 M Tris-HCl
pH 6.8 buffer and used for ELISA analysis and Western blotting of soluble APP cleavage products. The pellets were sonicated in RIPA buffer and
3. Kim et al. • TACE Inhibition Does Not Enhance A Production
12054 • J. Neurosci., November 12, 2008 • 28(46):12052–12061
centrifuged at 100,000 g for 1 h at 4°C. The
supernatants were saved and used for Western
blotting of flAPP and ␣-tubulin.
Cell and tissue lysates were electrophoresed
on 7.5% Tris-glycine acrylamide gels for flAPP,
sAPP␣/, and ␣-tubulin, and transferred to nitrocellulose. For CTFs (C-terminal fragments)
immunoblotting, samples were immunoprecipitated with 5685, a rabbit polyclonal antibody raised against a C-terminal peptide fraction of APP (Lee et al., 2005), before
electrophoresis on 16.5% Tris-tricine gels
(Bio-Rad). Full-length APP and CTFs were
probed with 5685. sAPP␣ was probed with 2B3,
an end-specific sAPP␣ monoclonal antibody
(IBL America). sAPP was specifically probed
with C5A4/2, a rabbit polyclonal antibody
raised against a synthetic peptide (CSEVKM)
corresponding to the C terminus of sAPP (Lee
et al., 2005). ␣-Tubulin was used as a loading
control for general protein contents. Immunobands were detected with species-specific
horseradish peroxidase-conjugated anti-IgG
antibodies (Santa Cruz Biotechnology), and developed with enhanced chemiluminescence
(ECL; PerkinElmer Life Sciences). ECL signal
was visualized and quantified with a LAS-3000
imager and MultiGauge version 2.3 software
(Fujifilm Life Science). Signal linearity was verified in a separate experiment (data not shown).
Sandwich ELISA analysis and alkaline phosphatase assay. To measure A1– 40 and
A1– 42 levels, both conditioned media and
RIPA lysates were assayed with A sandwich
ELISAs as previously described (Lee et al.,
2003). Briefly, BNT-77 (anti-A11–28) was
used to capture endogenous mouse A1– 40
and A1– 42 from wild-type mouse brains.
Ban50 (anti-A1–10) was used as a capturing
antibody for measuring both A1– 40 and
A1– 42 peptides for all other application.
Horseradish peroxidase (HRP)-conjugated
BA-27 and BC-05 were used to report A species ending at position 40 and 42. For quantification of A levels, synthetic A1– 40 and
A1– 42 (Bachem Bioscience) were serially diluted in corresponding buffer or media to generate standard curves. LN27, a monoclonal antibody that binds to the N-terminal 200 amino
acid residues of APP (Wertkin et al., 1993), was
used as capturing antibody for measuring
sAPP␣ in conditioned media. HRP-conjugated
Ban50 was used to report sAPP␣. C-terminal
tagged alkaline phosphatase activity in conditioned media was used as a surrogate marker for
TNF␣ secretion, using the p-Nitrophenyl phosphate liquid substrate system (Sigma).
Results
Figure 2. Inhibition of sAPP␣ by BMS-561392 does not coincide with an increase in -cleavage products of APP. A, Immunoblot of full-length APP and ␣-tubulin in cell lysates, and sAPP␣ and sAPP in conditioned media, from CHO-APPwt cell. RIPA
lysates (20 g) and 20 l of conditioned media were resolved in 7.5% Tris-glycine SDS-gels, blotted and probed with 5685
(fl-APP), Ban50 (sAPP␣), and ␣-tubulin antibodies. Whereas the level of sAPP␣ was reduced by BMS-561392 in a dosedependent manner, the sAPP level in the media remained unchanged. Consistent levels of fl-APP and tubulin confirmed a lack
of BMS-561392 toxicity. B, Immunoblot and quantitation of APP C-terminal fragments (CTFs). RIPA lysates (500 g) were
immunoprecipitated with 5685 antibody, resolved in a 16.5% Tris-tricine SDS-gel, blotted and probed with the same antibody.
Note that the levels of CTF, a pan-precursor protein of A, remain unchanged after BMS-561392 treatment. Quantification was
based on the means of three independent experiments ϮSE measurements. ϩp Ͻ 0.01, two-tailed Student’s t test; n.s.*, not
significant, two-tailed Student’s t test, p ϭ 0.150; n.s.**, not significant, one-way ANOVA, p ϭ 0.598. #Results are shown in
arbitrary unit (DMSO treatment as 1.0). C, A40 and 42 in conditioned media after BMS-561392 treatment. Quantification was
based on the mean of six independent experiments ϮSE measurements. n.s.*, Not significant, one-way ANOVA, p ϭ 0.902;
n.s.**, not significant, one-way ANOVA, p ϭ 0.874. #Results are normalized to fl-APP levels. D, Reduction of sAPP␣ secretion with
no concomitant change in A secretion in primary neurons derived from Tg2576 mice cultured for 7 d with or without BMS561392 for 24 h. RIPA lysates (20 g) were in 7.5% Tris-glycine SDS-gels, blotted, and probed with 5685 (fl-APP), Ban50 (sAPP␣),
C5A4/2 (sAPP), and ␣-tubulin antibodies. Quantification was based on the means of three independent experiments ϮSE
measurements. n.s.*, Not significant, one-way ANOVA, p ϭ 0.943; n.s.**, not significant, one-way ANOVA, p ϭ 0.996. #Results
are normalized to fl-APP levels.
BMS-561392 is a potent inhibitor of ␣cleavage of pro-TNF␣ and APP
To evaluate the efficacy of BMS-561392 in cell culture, different
concentrations of the compound were applied to CHO cells expressing the precursor, membrane-bound form of TNF␣ (proTNF␣). TAPI-I, a broad-range MMP/ADAM inhibitor, was also
included for comparison. After 24 h of treatment, we measured
the levels of soluble TNF␣, the product of TACE cleavage, in
culture media. Both inhibitors caused a dose-dependent inhibi-
tion of TNF␣ secretion (Fig. 1 A), with BMS-561392 being more
potent (50% inhibition with respect to DMSO controls was
achieved with 0.15 M BMS-561392 and 0.90 M TAPI-I).
To test the effect of BMS-561392 on the processing of APP,
another TACE substrate, CHO cells expressing either human
wild-type APP (APPwt) or human APP with the previously described Swedish mutation (APPswe) (Citron et al., 1992) were
treated with different concentrations of this compound for 24 h.
Secretion of sAPP␣ was reduced in a dose-dependent manner by
4. Kim et al. • TACE Inhibition Does Not Enhance A Production
J. Neurosci., November 12, 2008 • 28(46):12052–12061 • 12055
BMS-561392 as detected by a sAPP␣ sandwich ELISA (Fig. 1 B).
TAPI-I also caused a decrease in sAPP␣ secretion from CHOAPPwt cells, although with less potency relative to BMS-561392.
No cell toxicity was observed with any of the treatment conditions. Interestingly, BMS-561392 inhibits sAPP␣ secretion more
strongly in the CHO-APPswe cells than in CHO-APPwt cells, and
although we are unsure why there is this differential effect, these
data confirm that BMS-561392 is a potent inhibitor of TACE in
cell culture.
BMS-561392 does not change the level of -cleavage products
of APP
Because APP is also cleaved by BACE, inhibition of TACE may
increase the availability of the APP substrate for BACE cleavage,
thereby enhancing the production of the -secretase cleavage
products, including sAPP, CTF and 〈40/42. Unexpectedly,
BMS-561392 did not change the level of APP -cleavage products
in CHO-APPwt cells, including CTF and A40/42, whereas it
significantly reduced sAPP␣ secretion and CTF␣ levels (Fig. 2 A–
C). This de-coupling between ␣- and -cleavage of APP was also
observed in CHO-APPswe cells (data not shown). BMS-561392
also inhibited sAPP␣ secretion from primary cortical neurons
derived from Tg2576 mice, and similar to what was observed with
CHO cells, A40/42 secretion levels remained unchanged (Fig.
2 D). These results suggest that TACE inhibition does not lead to
greater APP processing by BACE.
BACE inhibition does not increase sAPP␣ secretion in APPexpressing CHO cells
Because TACE inhibition did not appear to affect BACE processing of APP, we asked whether conversely BACE inhibition might
alter TACE cleavage of APP. To test this, we treated CHO695wt
cells with increasing concentrations of the selective Merck BACE
inhibitor III. As shown in Figure 3, the secretion of sAPP and
A40/42 were drastically reduced by this BACE inhibitor,
whereas sAPP␣ secretion was unchanged. This result demonstrates that these enzymes do not compete with each other for the
processing of APPwt.
PMA-activated ␣-secretase activity results in decreased A
secretion that is normalized after BMS-561392 treatment
One explanation for the lack of coupling between TACE and
BACE cleavage of APP is that the enzymes “see” two separate
pools of substrate; therefore, manipulation of one pathway would
not affect the other. However, this hypothesis is inconsistent with
a previously published observation that TACE and BACE compete for APP substrate when TACE activity is potentiated by
phorbol 12-myristate 13-acetate (PMA)-treatment (Skovronsky
et al., 2000). As shown in Figure 4, we also observed a PMAinduced increase of ␣-secretase activity that resulted in enhanced
sAPP␣ secretion with a concomitant decrease in A40/42 secretion. BMS-561392 treatment in the PMA-activated condition effectively inhibited ␣-secretase and normalized A40 and A42
secretion to that seen in control cells.
The results obtained with PMA treatment appear to contradict our previous data which suggested that TACE and BACE do
not compete for APP. One way to reconcile both observations is
to hypothesize that the two enzymes reside in the same subcellular compartment, as has previously been suggested (Skovronsky
et al., 2000), but that the APP levels in the compartment are
normally not substrate limiting for TACE and BACE. However,
the combined TACE and BACE catalytic activity could exceed the
amount of available APP when TACE activity is significantly up-
Figure 3. BACE inhibitor does not increase sAPP␣ secretion from CHO695wt cells. A, Immunoblot of full-length APP and ␣-tubulin in cell lysates, and sAPP␣ and sAPP in conditioned
media, from CHO-APPwt cells. RIPA lysates (20 g) and 20 l of conditioned media were
resolved in 7.5% Tris-glycine SDS-gels, blotted and probed with 5685 (fl-APP), Ban50 (sAPP␣),
and ␣-tubulin antibodies. B, Quantification of sAPP␣, A40, and A42 levels in conditioned
media by sandwich ELISAs. BACE inhibitor potently decreases the secretion of A, but does not
induce higher sAPP␣ secretion. Quantification was based on the means of three independent
experiments ϮSE measurements. n.s., Not significant, one-way ANOVA, p ϭ 0.945. #Results
are normalized to fl-APP levels. sAPP␣ levels are shown in arbitrary units (average of DMSO
treatments as 1.0).
regulated by phorbol ester, thereby resulting in enzyme competition for substrate.
BACE inhibition increases sAPP␣ secretion from trans-Golgi
network-targeted APP
To further examine the hypothesis that TACE and BACE share a
common compartment, studies were conducted in which the
normal intracellular trafficking of APP was altered. CHO cells
were stably transfected with APP-TGN, a fusion construct in
which the entire cytoplasmic domain of APP is replaced with the
rat furin domain that enhances retention in the TGN (Chyung et
al., 1997; Skovronsky et al., 2000; Huse et al., 2002). Because the
5. Kim et al. • TACE Inhibition Does Not Enhance A Production
12056 • J. Neurosci., November 12, 2008 • 28(46):12052–12061
TGN has been proposed to be a site of ␣and -secretase action, increasing APP retention time in the TGN should result in
greater accessibility by these enzymes. As
shown in Figure 5, addition of the Merck
BACE inhibitor III to APP-TGN cells resulted in an increased production of
sAPP␣ relative to the vehicle-treated APPTGN cells, a result that is consistent with
the shared compartment hypothesis.
However, this data would appear to conflict with the results obtained with APPwt
cells (Fig. 3), where BACE inhibition did
not change sAPP␣ levels. One important
difference between the APP-TGN and APPwt cells is that the targeting sequence in
the former does not allow for normal APP
progression from the TGN outward to the
plasma membrane. Thus, whereas APP
concentration is unlikely to increase significantly within the TGN as a result of
BACE inhibition in APPwt cells because of
continued APP transit to the plasma membrane, APP would accumulate within the
TGN of APP-TGN cells. Accordingly, in
the absence of APP outflow, BACE inhibition would result in an even greater increase of TGN APP concentration that
would allow for increased sAPP␣ production by TACE.
Figure 4. BMS-561392 inhibits PMA-activated ␣-secretase activity, enhancing A secretion compared with PMA-only treatment. A, Immunoblot of full-length APP and ␣-tubulin in cell lysates, and sAPP␣ and sAPP in conditioned media, from
CHO-APPwt cells. B, Quantitation of sAPP␣ level in conditioned media, measured by densitometry of a 2B3 Western blot. C, A40
and A42 levels in conditioned media. PMA activates ␣-secretase activity and reduces A secretion, and BMS-561392 reverses
the effect of PMA on CHO695wt cells. Quantification based on the means of three independent experiments ϮSE measurements.
1
p Ͻ 0.05 ϳ 6p Ͻ 0.05; two-tailed Student’s t test. #Results are normalized to fl-APP levels. sAPP␣ levels are shown in arbitrary
units (average of DMSO only treatments as 1.0).
sAPP␣ levels were significantly reduced
in BMS-561392 infused Tg2576 mouse
hippocampi without an increase in
A levels
To test whether BMS-561392 acts as a potent TACE inhibitor in
vivo, we directly infused this compound or vehicle with osmotic
pumps into the right lateral ventricle of Tg2576 mice for 14 d.
Direct infusion was necessary because BMS-561392 does not
penetrate the blood– brain barrier [J. M. Trzaskos (Bristol-Myers
Squibb) personal communication]. As shown in Figure 6, A and
B, sAPP␣ levels in BMS-561392-infused mice were significantly
reduced in the hippocampus, but not in the cortex. This may be
because of the proximity of the hippocampus to the site of infusion, resulting in a greater concentration of BMS-561392 in interstitial fluid in the hippocampus than in the cortex, which is
much larger and farther away from the lateral ventricle. These
data show that BMS-561392 is an inhibitor of ␣-cleavage of APP
in vivo. Unlike BMS-561392, TAPI-I failed to lower sAPP␣ in the
hippocampus. This may reflect the higher potency of BMS561392 than TAPI-1 on TACE (Fig. 1). Although the inhibitory
effect of BMS-561392 on brain sAPP␣ levels appears to be somewhat less profound than what was observed in vitro (Fig. 1 B), it is
difficult to directly compare the cell-based studies with the more
complex in vivo model. Nonetheless, we note that the ventricular
infusion of BMS-561392 resulted in a highly significant 40% reduction in hippocampal sAPP␣, and it is possible that higher
doses of compound might have resulted in nearly complete
␣-secretase inhibition.
We also measured A40/42 levels in brain extracts and CSF of
Tg2576 mice. Similar to cell-based experiments, A40/42 levels
were not statistically different in vehicle- versus BMS-561392infused mice (Fig. 6C). Thus, the in vivo result agrees with our
observation in cultured cells that ␣- and -secretase do not compete for APP under normal conditions. Finally, an analysis of
full-length APP did not reveal any significant differences among
the treatment groups (data not shown).
sAPP␣ levels were significantly reduced in BMS-561392infused wild-type mice without an increase in A levels
Although A levels remained unchanged in BMS-561392infused Tg2576 mice, the experimental system may not reflect the
effect of TACE inhibition on A generation in human brains
where APP is not artificially over-expressed. We therefore determined whether BMS-561392 affects A secretion in wild-type
mice expressing only endogenous APP. To detect the low level of
A and other APP cleavage products, we sequentially extracted
wild-type mouse brains with DEA-NaCl and RIPA buffers
(Miller et al., 2003). DEA extraction effectively separated sAPP␣
and sAPP from fl-APP (supplemental Fig. 2, available at www.
jneurosci.org as supplemental material). Figure 7, A and B, show
that sAPP␣ levels were reduced in the brains of BMS-561392
infused non-transgenic wild type mice. Unlike Tg2576 mice,
sAPP␣ was significantly lower in the cortices as well as the hippocampi of the drug-infused brains compared with vehicletreated brains. One possible explanation is that the lower level of
APP in the cortices of wild-type mice relative to Tg2576 mice
enabled more effective inhibition of TACE cleavage of APP by
BMS-561392. Whereas BMS-561392 inhibited sAPP␣ generation
in wild-type mouse brains, A levels remained unchanged in
both brain lysates and cerebrospinal fluids (Fig. 7C). Similarly,
full-length APP levels were not statistically different between
6. Kim et al. • TACE Inhibition Does Not Enhance A Production
Figure 5. BACE inhibitor increases sAPP␣ secretion from CHO cells expressing TGN-targeting
APP. A, Immunoblot of full-length APP and ␣-tubulin in cell lysates, and sAPP␣ and sAPP in
conditioned media, from CHO-APPwt and CHO-APP-TGN cells. APP-TGN migrated slower because of the addition of the furin sequence (Chyung et al., 1997; Skovronsky et al., 2000; Huse et
al., 2002). RIPA lysates (20 g) and 20 l of conditioned media were resolved in 7.5% Trisglycine SDS-gels, blotted and probed with 5685 (fl-APP), Ban50 (sAPP␣), and ␣-tubulin antibodies. B, Quantification of sAPP␣ in conditioned media. Unlike CHO-APPwt cells, BACE inhibitor increases sAPP␣ secretion from CHO-APP-TGN cells. Quantification are means of three
independent experiments ϮSE measurements. n.s., Not significant, two-tailed Student’s t test,
p ϭ 0.210; p Ͻ 0.005, paired two-tailed Student’s t test. #Results are normalized to fl-APP
levels in each cell line. sAPP␣ levels are shown in arbitrary units (DMSO treatment as 1.0).
control and drug-treated mouse brains (data not shown). These
data demonstrate that BMS-561392 does not enhance A secretion either in wild-type mice or in transgenic mice that overexpress APP. Thus, anti-inflammatory drugs targeting TACE
may not necessarily promote an increase in A production.
Discussion
Although previous studies suggested that suppression of glial activation and a reduction in neuroinflammation might prove beneficial in AD (McGeer et al., 2006), the effects of selective TNF␣
reduction remain unknown because of a lack of agents that can
specifically suppress the biosynthesis or release of this inflammatory cytokine in the brain. In this study, we demonstrated that a
highly selective TACE inhibitor, BMS-561392, reduces TNF␣ se-
J. Neurosci., November 12, 2008 • 28(46):12052–12061 • 12057
cretion in cultured cells and also inhibits the cleavage of another
TACE substrate, APP, both in cell culture and in mouse models.
It should be noted that TACE cleavage of APP precludes the
generation of potentially harmful A peptides, and thus, inhibition of TACE might be predicted to lead to elevated A levels that
could offset some or all of the possible therapeutic benefits of
TNF␣ suppression (Buxbaum et al., 1998; Lammich et al., 1999).
Our results show that BMS-561392 does not elevate A secretion
in cell culture at a dose that causes Ͼ50% reduction in TNF␣
secretion. The level of CTF, the precursor that is cleaved by
␥-secretase to yield A, also remained unchanged in BMS561392-treated cells, confirming that TACE inhibition did not
affect BACE processing of APP. We observed a similar lack of
effect of TACE inhibition on BACE cleavage of APP in an APPover-expression mouse model as well as in wild-type mice that
express only endogenous APP. Whereas sAPP␣ levels in the hippocampus were significantly reduced in BMS-561392-infused
mice, steady-state A levels remained the same. Similarly, CSF
A levels also showed no change after TACE inhibition. The lack
of correlation between ␣- and -cleavage of APP, both in vitro
and in vivo, suggests that there is little competition for APP between TACE and BACE. Thus, although BMS-561392 itself is not
suitable for the treatment of neuroinflammatory conditions because it does not readily cross the blood– brain barrier, these data
suggest that a brain-penetrant TACE inhibitor would be unlikely
to induce an undesirable elevation of brain A and therefore may
have potential as an AD therapeutic.
Although it appears that TACE and BACE do not normally
compete for APP, conditions can be created in which inhibition
of one of these enzymes can affect the amount of APP processed
by the other. For example, TACE inhibition by BMS-561392
caused an increased production of A peptides relative to untreated cells when the catalytic activity of TACE was significantly
enhanced by phorbol ester. Moreover, BACE inhibition in cells
that expressed a TGN-retained form of APP resulted in an elevation of sAPP␣. These data suggest that TACE and BACE are
colocalized within the TGN and that manipulation of the TGN
milieu can yield conditions whereby modulation of one secretase
can affect APP cleavage by the other. The steady-state level of APP
that transiently resides in the TGN during its trafficking to the
plasma membrane might normally be in excess of the combined
TACE and BACE catalytic activities, but the elevation of TACE
activity by phorbol ester may cause enough additional APP cleavage to make the TGN pool of APP limiting. On the other hand,
artificially retaining APP within the TGN via a targeting sequence
disrupts its normal outward flux such that BACE inhibition
would result in an elevation of TGN APP concentration, with a
consequent increase of sAPP␣ production by TACE.
Our observation of limited competition between TACE and
BACE for APP only under certain conditions, such as PKC activation, agrees with certain previous studies (Gabuzda et al., 1993;
Hung et al., 1993; Jacobsen et al., 1994; Blacker et al., 2002; Gandhi et al., 2004). However, there are other reports that would
appear to be in disagreement with our data in regard to whether
and under what conditions these two enzymes compete for APP.
(Dyrks et al., 1994; Buxbaum et al., 1998; LeBlanc et al., 1998). We
suggest that the differences may be attributable to our use of a
highly specific TACE inhibitor instead of a pleiotropic inhibitor
of the MMP/ADAM family and perhaps to cell-type specific behavior, especially with respect to the response toward PMA treatment. It should be noted, however, that we consistently demonstrated an absence of A enhancement by BMS-561392 in
7. 12058 • J. Neurosci., November 12, 2008 • 28(46):12052–12061
Kim et al. • TACE Inhibition Does Not Enhance A Production
Figure 6. BMS-561392 reduces the level of sAPP␣ in Tg2576 transgenic mouse hippocampi without changes in A level. A, BMS-561392 reduces sAPP␣ level in Tg2576 mouse hippocampi, but
TAPI-I fails to do so. RIPA lysates (20 g) of drug-treated mouse cortices and hippocampi were resolved in 7.5% Tris-glycine SDS-gels, blotted and probed with 5685 (fl-APP), 2B3 (sAPP␣), and
␣-tubulin antibodies. B, Quantification based on the average of n ϭ 5 for each group in six independent immunoblots ϮSE measurements. n.s. a, Not significant, one-way ANOVA, p ϭ 0.734; n.s. b,
not significant, unpaired two-tailed Student’s t test, p ϭ 0.236; ϩp Ͻ 0.0001; unpaired two-tailed Student’s t test. #Results are normalized to fl-APP levels and shown in arbitrary units (control
as 1.0). C, A40 and A42 levels in brain lysates and CSF showed no statistically significant change. n.s. 1 ϳ n.s. 6, Not significant, one-way ANOVA, p ϭ 0.735, 0.590, 0.647, 0.480, 0.550, and 0.606,
respectively.
cultured CHO cells and primary neurons, as well as in human
APP-expressing transgenic and wild-type mice.
Despite its upregulation in AD and other brain diseases (Fillit
et al., 1991; Mogi et al., 1994; Botchkina et al., 1997), the effects of
TNF␣ remain somewhat controversial because proinflammatory
cytokines do not usually cause death of neurons. Moreover, it has
been suggested that TNF␣ can act as a neuroprotective agent by
suppressing CDK5 hyperactivity (Orellana et al., 2007). However, identification of single nucleotide polymorphisms in the
TNF␣ gene that are linked to AD strongly supports the hypothesis that TNF␣ is a significant contributing factor to neuronal
degeneration (Collins et al., 2000; Lio et al., 2006; Ramos et al.,
2006). A G-to-A mutation in the promoter region at position
Ϫ308 of the TNF␣ gene has been shown to be associated with
higher intrathecal level of this cytokine and earlier onset of Alzheimer’s disease. In contrast, an allele at position Ϫ863 of the
TNF␣ gene locus results in lower serum TNF␣ levels and reduced
risk of AD. Interestingly, the presence of the APOE 4 allele, a
major risk factor for late-onset AD, is synergetic to the effects of
the TNF␣ position Ϫ308 haplotype in lowering the age of onset
of the disease.
The prominent brain inflammation and neurodegeneration
that is seen in transgenic mice over-expressing TNF␣ or its receptors indicates that TNF␣ signaling cascades can lead to significant
neuronal loss (Probert et al., 1995; Akassoglou et al., 1997; Stalder
et al., 1998). TNF␣ secretion from activated microglial can be
induced by neurotoxic insults, including A40 and 42 peptides
(Klegeris et al., 1997; Fiala et al., 1998; Yates et al., 2000), and
TNF␣ can enhance the secretion of A. Thus the interplay between A and TNF␣ could lead to a vicious and damaging cycle
in the AD brain (Blasko et al., 1999). Interestingly, deletion of
TNF type I receptor (TNFR1, p55) gene in APP23 transgenic
mice diminished amyloid plaque formation and improved cognitive deficits by downregulating BACE1 promoter activity (He et
al., 2007). Infliximab and thalidomide, a TNF␣ antagonist and an
inhibitor of TNF␣ production, were shown to prevent Amediated LTP inhibition in mice, suggesting cross-interaction
between a TNF␣ signaling cascade and A toxicity (Wang et al.,
2005). TNF␣ can also stimulate other inflammatory pathways,
including those mediated by NF-〉 and cyclooxygenase (COX)
2 (Meichle et al., 1990; Geng et al., 1995; Perry et al., 2001), of
which inhibitors have been suggested to lower the risk of AD
(Lim et al., 2000; Sung et al., 2004; Heneka et al., 2005). Thus,
TNF␣ may cause neuronal damage and memory loss through
multiple mechanisms (Tarkowski et al., 1999; Sastre et al., 2006).
Although a phase II trial of BMS-561392 for rheumatoid arthritis was halted because of signs of mild hepatotoxicity, several
other anti-TNF␣ agents, such as infliximab, are widely used for
the treatment of autoimmune diseases. Interestingly, curcumin, a
natural product that reduces amyloid accumulation in mouse
8. Kim et al. • TACE Inhibition Does Not Enhance A Production
J. Neurosci., November 12, 2008 • 28(46):12052–12061 • 12059
Figure 7. BMS-561392 reduces the level of sAPP␣ in B6/SJL F1 wild-type mouse brains without changes in A level. A, BMS-561392 reduces sAPP␣ level in drug-treated SJL/B6 F1 mouse
cortices and hippocampi, whereas A40 and A42 levels in brain lysates and CSF were unchanged. Fifty g of DEA/NaCl lysates (for sAPP␣) and RIPA lysates (for fl-APP and ␣-tubulin) of
drug-treated mouse cortices and hippocampi were resolved in 7.5% Tris-glycine SDS-gels, blotted and probed with 5685 (fl-APP), 2B3 (sAPP␣), and ␣-tubulin antibodies. B, Quantification based
on the average of n ϭ 6 for each group ϮSE measurements. ϩp Ͻ 0.05; 2ϩp Ͻ 0.05, unpaired two-tailed Student’s t test. #Results are normalized to fl-APP levels and shown in arbitrary units
(control as 1.0). C, A40 and A42 levels in brain lysates and CSF were unchanged. n.s. 1 ϳ n.s. 6, Not significant, unpaired two-tailed Student’s t test, p ϭ 0.877, 0.482, 0.633, 0.574, 0.928, and
0.785, respectively.
models of AD (Yang et al., 2005; Garcia-Alloza et al., 2007), is an
anti-inflammatory compound that lowers levels of serum TNF␣
in a rat model of acute pancreatitis (Gulcubuk et al., 2006). Despite the lack of extensive evidence for the therapeutic effects of
TACE-targeted anti-TNF␣ treatment in AD, this approach may
be well worth pursuing. However, it should be recognized that
one consequence of TACE inhibition will be a reduction of
sAPP␣, which has been proposed to have neurotrophic properties in cultured neurons and in a rat traumatic brain injury model
(Mattson et al., 1993; Yamamoto et al., 1994; Thornton et al.,
2006). Although there is no evidence showing that a reduction of
sAPP␣ by TACE inhibition is detrimental, this possibility deserves further exploration in future studies.
Another consequence of TACE inhibition is that any positive
effects of TNF␣ on neuronal systems will be compromised. For
example, it has been suggested that TNF␣ may protect neurons
against cellular stress and is even neurotrophic through TNF receptor II (TNFR2) (Cheng et al., 1994; Yang et al., 2002). However, it is unclear that inhibition of TNF␣ secretion in adult brain
will have negative consequences that will outweigh the likely positive effects of preventing TNF␣-mediated inflammation.
In conclusion, we have demonstrated that the potent TACE
inhibitor, BMS-561392, does not induce or increase A secretion
under normal conditions. Therefore, although TACE and BACE
appear to reside in a shared TGN compartment, the use of TACE-
targeted anti-inflammatory drugs in AD may not be associated
with higher A generation in brain. Although BMS-561392 itself
is unlikely to be a candidate drug for AD because of its poor brain
penetration, the identification and development of improved
TACE inhibitors that gain access to the brain may be of be potential value in reducing the glial-mediated inflammation that has
been suggested to contribute to AD neuropathology.
References
Akassoglou K, Probert L, Kontogeorgos G, Kollias G (1997) Astrocytespecific but not neuron-specific transmembrane TNF triggers inflammation and degeneration in the central nervous system of transgenic mice.
J Immunol 158:438 – 445.
Akiyama H, Barger S, Barnum S, Bradt B, Bauer J, Cole GM, Cooper NR,
Eikelenboom P, Emmerling M, Fiebich BL, Finch CE, Frautschy S, Griffin
WS, Hampel H, Hull M, Landreth G, Lue L, Mrak R, Mackenzie IR,
McGeer PL, et al. (2000) Inflammation and Alzheimer’s disease. Neurobiol Aging 21:383– 421.
Blacker M, Noe MC, Carty TJ, Goodyer CG, LeBlanc AC (2002) Effect of
tumor necrosis factor-alpha converting enzyme (TACE) and metalloprotease inhibitor on amyloid precursor protein metabolism in human neurons. J Neurochem 83:1349 –1357.
Blasko I, Marx F, Steiner E, Hartmann T, Grubeck-Loebenstein B (1999)
TNFalpha plus IFNgamma induce the production of Alzheimer betaamyloid peptides and decrease the secretion of APPs. FASEB J 13:63– 68.
Botchkina GI, Meistrell ME 3rd, Botchkina IL, Tracey KJ (1997) Expression
of TNF and TNF receptors (p55 and p75) in the rat brain after focal
cerebral ischemia. Mol Med 3:765–781.
9. 12060 • J. Neurosci., November 12, 2008 • 28(46):12052–12061
Buxbaum JD, Liu KN, Luo Y, Slack JL, Stocking KL, Peschon JJ, Johnson RS,
Castner BJ, Cerretti DP, Black RA (1998) Evidence that tumor necrosis
factor alpha converting enzyme is involved in regulated alpha-secretase
cleavage of the Alzheimer amyloid protein precursor. J Biol Chem
273:27765–27767.
Cheng B, Christakos S, Mattson MP (1994) Tumor necrosis factors protect
neurons against metabolic-excitotoxic insults and promote maintenance
of calcium homeostasis. Neuron 12:139 –153.
Chesneau V, Becherer JD, Zheng Y, Erdjument-Bromage H, Tempst P, Blobel
CP (2003) Catalytic properties of ADAM19. J Biol Chem
278:22331–22340.
Chyung AS, Greenberg BD, Cook DG, Doms RW, Lee VM-Y (1997) Novel
beta-secretase cleavage of beta-amyloid precursor protein in the endoplasmic reticulum/intermediate compartment of NT2N cells. J Cell Biol
138:671– 680.
Citron M, Oltersdorf T, Haass C, McConlogue L, Hung AY, Seubert P, VigoPelfrey C, Lieberburg I, Selkoe DJ (1992) Mutation of the beta-amyloid
precursor protein in familial Alzheimer’s disease increases beta-protein
production. Nature 360:672– 674.
Collins JS, Perry RT, Watson B Jr, Harrell LE, Acton RT, Blacker D, Albert
MS, Tanzi RE, Bassett SS, McInnis MG, Campbell RD, Go RC (2000)
Association of a haplotype for tumor necrosis factor in siblings with lateonset Alzheimer disease: the NIMH Alzheimer Disease Genetics Initiative. Am J Med Genet 96:823– 830.
De Strooper B, Annaert W (2000) Proteolytic processing and cell biological
functions of the amyloid precursor protein. J Cell Sci 113:1857–1870.
Dominguez D, Tournoy J, Hartmann D, Huth T, Cryns K, Deforce S, Serneels
L, Camacho IE, Marjaux E, Craessaerts K, Roebroek AJ, Schwake M,
D’Hooge R, Bach P, Kalinke U, Moechars D, Alzheimer C, Reiss K, Saftig
P, De Strooper B (2005) Phenotypic and biochemical analyses of B.
J Biol Chem 280:30797–30806.
Dyrks T, Monning U, Beyreuther K, Turner J (1994) Amyloid precursor
¨
protein secretion and beta A4 amyloid generation are not mutually exclusive. FEBS Lett 349:210 –214.
Fiala M, Zhang L, Gan X, Sherry B, Taub D, Graves MC, Hama S, Way D,
Weinand M, Witte M, Lorton D, Kuo YM, Roher AE (1998) Amyloidbeta induces chemokine secretion and monocyte migration across a human blood– brain barrier model. Mol Med 4:480 – 489.
Fillit H, Ding WH, Buee L, Kalman J, Altstiel L, Lawlor B, Wolf-Klein G
(1991) Elevated circulating tumor necrosis factor levels in Alzheimer’s
disease. Neurosci Lett 129:318 –320.
Forman MS, Cook DG, Leight S, Doms RW, Lee VM-Y (1997) Differential
effects of the Swedish mutant amyloid precursor protein on beta-amyloid
accumulation and secretion in neurons and nonneuronal cells. J Biol
Chem 272:32247–32253.
Gabuzda D, Busciglio J, Yankner BA (1993) Inhibition of beta-amyloid production by activation of protein kinase C. J Neurochem 61:2326 –2329.
Gandhi S, Refolo LM, Sambamurti K (2004) Amyloid precursor protein
compartmentalization restricts beta-amyloid production: therapeutic
targets based on BACE compartmentalization. J Mol Neurosci
24:137–143.
Garcia-Alloza M, Borrelli LA, Rozkalne A, Hyman BT, Bacskai BJ (2007)
Curcumin labels amyloid pathology in vivo, disrupts existing plaques,
and partially restores distorted neurites in an Alzheimer mouse model.
J Neurochem 102:1095–1104.
Geng Y, Blanco FJ, Cornelisson M, Lotz M (1995) Regulation of
cyclooxygenase-2 expression in normal human articular chondrocytes.
J Immunol 155:796 – 801.
Gulcubuk A, Altunatmaz K, Sonmez K, Haktanir-Yatkin D, Uzun H, Gurel A,
Aydin S (2006) Effects of curcumin on tumour necrosis factor-alpha
and interleukin-6 in the late phase of experimental acute pancreatitis. J
Vet Med A Physiol Pathol Clin Med 53:49 –54.
He P, Zhong Z, Lindholm K, Berning L, Lee W, Lemere C, Staufenbiel M, Li
R, Shen Y (2007) Deletion of tumor necrosis factor death receptor inhibits amyloid beta generation and prevents learning and memory deficits
in Alzheimer’s mice. J Cell Biol 178:829 – 841.
Heneka MT, Sastre M, Dumitrescu-Ozimek L, Hanke A, Dewachter I, Kuiperi
C, O’Banion K, Klockgether T, Van Leuven F, Landreth GE (2005)
Acute treatment with the PPARgamma agonist pioglitazone and ibuprofen reduces glial inflammation and Abeta1– 42 levels in APPV717I transgenic mice. Brain 128:1442–1453.
Hung AY, Haass C, Nitsch RM, Qiu WQ, Citron M, Wurtman RJ, Growdon
Kim et al. • TACE Inhibition Does Not Enhance A Production
JH, Selkoe DJ (1993) Activation of protein kinase C inhibits cellular
production of the amyloid beta-protein. J Biol Chem 268:22959 –22962.
Huse JT, Liu K, Pijak DS, Carlin D, Lee VM-Y, Doms RW (2002) Betasecretase processing in the trans-Golgi network preferentially generates
truncated amyloid species that accumulate in Alzheimer’s disease brain.
J Biol Chem 277:16278 –16284.
Itai T, Tanaka M, Nagata S (2001) Processing of tumor necrosis factor by the
membrane-bound TNF-alpha-converting enzyme, but not its truncated
soluble form. Eur J Biochem 268:2074 –2082.
Jacobsen JS, Spruyt MA, Brown AM, Sahasrabudhe SR, Blume AJ, Vitek MP,
Muenkel HA, Sonnenberg-Reines J (1994) The release of Alzheimer’s
disease beta amyloid peptide is reduced by phorbol treatment. J Biol
Chem 269:8376 – 8382.
Klegeris A, Walker DG, McGeer PL (1997) Interaction of Alzheimer betaamyloid peptide with the human monocytic cell line THP-1 results in a
protein kinase C-dependent secretion of tumor necrosis factor-alpha.
Brain Res 747:114 –121.
Koike H, Tomioka S, Sorimachi H, Saido TC, Maruyama K, Okuyama A,
Fujisawa-Sehara A, Ohno S, Suzuki K, Ishiura S (1999) Membraneanchored metalloprotease MDC9 has an alpha-secretase activity responsible for processing the amyloid precursor protein. Biochem J
343:371–375.
Lammich S, Kojro E, Postina R, Gilbert S, Pfeiffer R, Jasionowski M, Haass C,
Fahrenholz F (1999) Constitutive and regulated alpha-secretase cleavage of Alzheimer’s amyloid precursor protein by a disintegrin metalloprotease. Proc Natl Acad Sci U S A 96:3922–3927.
LeBlanc AC, Koutroumanis M, Goodyer CG (1998) Protein kinase C activation increases release of secreted amyloid precursor protein without
decreasing A production in human primary neuron cultures. J Neurosci
18:2907–2913.
Lee EB, Skovronsky DM, Abtahian F, Doms RW, Lee VM-Y (2003) Secretion and intracellular generation of truncated Abeta in beta-site amyloidbeta precursor protein-cleaving enzyme expressing human neurons.
J Biol Chem 278:4458 – 4466.
Lee EB, Zhang B, Liu K, Greenbaum EA, Doms RW, Trojanowski JQ, Lee
VM-Y (2005) BACE overexpression alters the subcellular processing of
APP and inhibits Abeta deposition in vivo. J Cell Biol 168:291–302.
Lim GP, Yang F, Chu T, Chen P, Beech W, Teter B, Tran T, Ubeda O, Ashe
KH, Frautschy SA, Cole GM (2000) Ibuprofen suppresses plaque pathology and inflammation in a mouse model for Alzheimer’s disease.
J Neurosci 20:5709 –5714.
Lio D, Annoni G, Licastro F, Crivello A, Forte GI, Scola L, Colonna-Romano
G, Candore G, Arosio B, Galimberti L, Vergani C, Caruso C (2006) Tumor necrosis factor-alpha -308A/G polymorphism is associated with age
at onset of Alzheimer’s disease. Mech Ageing Dev 127:567–571.
Mattson MP, Cheng B, Culwell AR, Esch FS, Lieberburg I, Rydel RE (1993)
Evidence for excitoprotective and intraneuronal calcium-regulating roles
for secreted forms of the beta-amyloid precursor protein. Neuron
10:243–254.
McGeer PL, McGeer EG (2001) Polymorphisms in inflammatory genes and
the risk of Alzheimer disease. Arch Neurol 58:1790 –1792.
McGeer PL, Rogers J, McGeer EG (2006) Inflammation, anti-inflammatory
agents and Alzheimer disease: the last 12 years. J Alzheimers Dis
9:271–276.
Meichle A, Schutze S, Hensel G, Brunsing D, Kronke M (1990) Protein
¨
¨
kinase C-independent activation of nuclear factor kappa B by tumor necrosis factor. J Biol Chem 265:8339 – 8343.
Miller BC, Eckman EA, Sambamurti K, Dobbs N, Chow KM, Eckman CB,
Hersh LB, Thiele DL (2003) Amyloid-beta peptide levels in brain are
inversely correlated with insulysin activity levels in vivo. Proc Natl Acad
Sci USA 100:6221– 6226.
Mogi M, Harada M, Riederer P, Narabayashi H, Fujita K, Nagatsu T (1994)
Tumor necrosis factor-alpha (TNF-alpha) increases both in the brain and
in the cerebrospinal fluid from parkinsonian patients. Neurosci Lett
165:208 –210.
Orellana DI, Quintanilla RA, Maccioni RB (2007) Neuroprotective effect of
TNFalpha against the beta-amyloid neurotoxicity mediated by CDK5 kinase. Biochim Biophys Acta 1773:254 –263.
Perry RT, Collins JS, Wiener H, Acton R, Go RC (2001) The role of TNF and
its receptors in Alzheimer’s disease. Neurobiol Aging 22:873– 883.
Probert L, Akassoglou K, Pasparakis M, Kontogeorgos G, Kollias G (1995)
Spontaneous inflammatory demyelinating disease in transgenic mice
10. Kim et al. • TACE Inhibition Does Not Enhance A Production
showing central nervous system-specific expression of tumor necrosis
factor alpha. Proc Natl Acad Sci U S A 92:11294 –11298.
Ramos EM, Lin MT, Larson EB, Maezawa I, Tseng LH, Edwards KL, Schellenberg GD, Hansen JA, Kukull WA, Jin LW (2006) Tumor necrosis
factor alpha and interleukin 10 promoter region polymorphisms and risk
of late-onset Alzheimer disease. Arch Neurol 63:1165–1169.
Sastre M, Klockgether T, Heneka MT (2006) Contribution of inflammatory
processes to Alzheimer’s disease: molecular mechanisms. Int J Dev Neurosci 24:167–176.
Skovronsky DM, Moore DB, Milla ME, Doms RW, Lee VM (2000) Protein
kinase C-dependent alpha-secretase competes with beta-secretase for
cleavage of amyloid-beta precursor protein in the trans-golgi network.
J Biol Chem 275:2568 –2575.
Stalder AK, Carson MJ, Pagenstecher A, Asensio VC, Kincaid C, Benedict M,
Powell HC, Masliah E, Campbell IL (1998) Late-onset chronic inflammatory encephalopathy in immune-competent and severe combined
immune-deficient (SCID) mice with astrocyte-targeted expression of tumor necrosis factor. Am J Pathol 153:767–783.
Sung S, Yang H, Uryu K, Lee EB, Zhao L, Shineman D, Trojanowski JQ, Lee
VM, Pratico D (2004) Modulation of nuclear factor-kappa B activity by
`
indomethacin influences A beta levels but not A beta precursor protein
metabolism in a model of Alzheimer’s disease. Am J Pathol
165:2197–2206.
Tarkowski E, Blennow K, Wallin A, Tarkowski A (1999) Intracerebral production of tumor necrosis factor-alpha, a local neuroprotective agent, in
Alzheimer disease and vascular dementia. J Clin Immunol 19:223–230.
Thornton E, Vink R, Blumbergs PC, Van Den Heuvel C (2006) Soluble
amyloid precursor protein alpha reduces neuronal injury and improves
functional outcome following diffuse traumatic brain injury in rats. Brain
Res 1094:38 – 46.
J. Neurosci., November 12, 2008 • 28(46):12052–12061 • 12061
Wang Q, Wu J, Rowan MJ, Anwyl R (2005) Beta-amyloid inhibition of
long-term potentiation is mediated via tumor necrosis factor. Eur J Neurosci 22:2827–2832.
Wertkin AM, Turner RS, Pleasure SJ, Golde TE, Younkin SG, Trojanowski
JQ, Lee VM-Y (1993) Human neurons derived from a teratocarcinoma
cell line express solely the 695-amino acid amyloid precursor protein and
produce intracellular beta-amyloid or A4 peptides. Proc Natl Acad Sci
U S A 90:9513–9517.
Wolf BA, Wertkin AM, Jolly YC, Yasuda RP, Wolfe BB, Konrad RJ, Manning
D, Ravi S, Williamson JR, Lee VM (1995) Muscarinic regulation of Alzheimer’s disease amyloid precursor protein secretion and amyloid betaprotein production in human neuronal NT2N cells. J Biol Chem
270:4916 – 4922.
Yamamoto K, Miyoshi T, Yae T, Kawashima K, Araki H, Hanada K, Otero
DA, Roch JM, Saitoh T (1994) The survival of rat cerebral cortical neurons in the presence of trophic APP peptides. J Neurobiol 25:585–594.
Yang F, Lim GP, Begum AN, Ubeda OJ, Simmons MR, Ambegaokar SS, Chen
PP, Kayed R, Glabe CG, Frautschy SA, Cole GM (2005) Curcumin inhibits formation of amyloid beta oligomers and fibrils, binds plaques, and
reduces amyloid in vivo. J Biol Chem 280:5892–5901.
Yang L, Lindholm K, Konishi Y, Li R, Shen Y (2002) Target depletion of
distinct tumor necrosis factor receptor subtypes reveals hippocampal
neuron death and survival through different signal transduction pathways. J Neurosci 22:3025–3032.
Yates SL, Burgess LH, Kocsis-Angle J, Antal JM, Dority MD, Embury PB,
Piotrkowski AM, Brunden KR (2000) Amyloid beta and amylin
fibrils induce increases in proinflammatory cytokine and chemokine
production by THP-1 cells and murine microglia. J Neurochem 74:
1017–1025.