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JUN jun proto- oncogene [Homo sapiens (human)] - Gene. AGE- RAGE signaling pathway in diabetic complications, organism- specific biosystem (from KEGG). AGE- RAGE signaling pathway in diabetic complications, organism- specific biosystem. Advanced glycation end products (AGEs) are a complex group of compounds produced through the non- enzymatic glycation and oxidation of proteins, lipids and nucleic acids, primarily due to aging and un.. Inversely Correlated in Systemic Anaplastic Large Cell Lymphoma1. c-Jun activation domain binding protein-1 (JAB1). dine; ALCL, anaplastic large cell lymphoma; JAB1, c-Jun activation binding protein 1. Vol. 9. The positive feedback system using a fusion protein of the sequence-specific DNA binding domain of yeast GAL4 and the transcription activation domain. sterol regulatory element-binding protein-1. Stimulation of c-Jun. Jun activation domain binding protein, activator protein 1, enhancer-binding. Results suggest that VEGF induced by hyperbaric oxygen is through c-Jun/AP-1 activation, and through simultaneous activation of ERK and JNK. 1. Mol Cancer Res. 2007 Oct;5(10):1063-72. Hepatitis B virus pre-S2 mutant surface antigen induces degradation of cyclin-dependent kinase inhibitor p27Kip1 through c-Jun activation domain-binding protein 1. Hsieh YH(1), Su IJ. Abstract. S100A7 is among the most highly expressed genes in preinvasive breast cancer, is a marker of poor survival when expressed in invasive disease, and promotes breast tumor progression in experimental models. C-jun represses p53 transcription by binding to a variant AP-1 site in the. The cyclic change of the c-jun protein levels is significant in the proliferation and apoptosis. AP-1. c-Fos; c-Jun; c-Myc; Mitochondrion. Jun Activation Domain Binding Protein 3; Enhancer-Binding. Protein details for JUN Gene (UniProtKB/Swiss-Prot). c-Jun (B-1) p-c-Jun (Tyr 170) p-c-Jun (Thr 93). Sigma-Aldrich offers abstracts and full-text articles by Ethan D Emberley, Yulian Niu, Linda Curtis, Sandra Troup, Sanat K Mandal, Jeffery N Myers, Spencer B Gibson, Leigh C Murphy, Peter H Watson. The S100A7-c-Jun Activation Domain Binding Protein 1 Pathway Enhances Prosurvival Pathways in Breast Cancer Ethan D. Emberley,1,3 Yulian Niu, 2,3Linda Curtis, Sandra Troup, Sanat K. Mandal,4 Jeffery N. Myers,5 Spencer B. ![]() AGE- RAGE signaling pathway in diabetic complications, conserved biosystem (from KEGG). AGE- RAGE signaling pathway in diabetic complications, conserved biosystem. Advanced glycation end products (AGEs) are a complex group of compounds produced through the non- enzymatic glycation and oxidation of proteins, lipids and nucleic acids, primarily due to aging and un.. AGE/RAGE pathway, organism- specific biosystem (from Wiki. Pathways). AGE/RAGE pathway, organism- specific biosystem. Advanced glycation end products (AGEs) are heterogeneous group of non- enzymatic malliard reaction products of aldose sugar with proteins and lipids. Formation of AGEs is an indicator of one of the ma.. ATF- 2 transcription factor network, organism- specific biosystem (from Pathway Interaction Database). ATF- 2 transcription factor network, organism- specific biosystem. ATF- 2 transcription factor network. Activated TLR4 signalling, organism- specific biosystem (from REACTOME). Activated TLR4 signalling, organism- specific biosystem. TLR4 is unique among the TLR family in its ability to recruit four adapters to activate two distinct signaling pathways. One pathway is activated by the pair of the adapters Mal or TIRAP (Toll/inter.. Activation of HOX genes during differentiation, organism- specific biosystem (from REACTOME). Activation of HOX genes during differentiation, organism- specific biosystem. Hox genes encode proteins that contain the DNA- binding homeobox motif and control early patterning of segments in the embryo as well as later events in development (reviewed in Rezsohazy et al. Activation of anterior HOX genes in hindbrain development during early embryogenesis, organism- specific biosystem (from REACTOME). Activation of anterior HOX genes in hindbrain development during early embryogenesis, organism- specific biosystem. In mammals, anterior Hox genes may be defined as paralog groups 1 to 4 (Natale et al. Activation of the AP- 1 family of transcription factors, organism- specific biosystem (from REACTOME). Activation of the AP- 1 family of transcription factors, organism- specific biosystem. AP-1 transcription factor is. Apart from the “leucine zipper” and the “basic region” which are important for dimerization and DNA-binding, the c-jun protein. The outcome of AP-1 activation is dependent.Activator protein- 1 (AP- 1) is a collective term referring to a group of transcription factors that bind to promoters of target genes in a sequence- specific manner. AP- 1 family consists of hetero- and.. Ah. R pathway, organism- specific biosystem (from Wiki. Pathways). Ah. R pathway, organism- specific biosystem. Ah. R pathway. Amphetamine addiction, organism- specific biosystem (from KEGG). Amphetamine addiction, organism- specific biosystem. Amphetamine is a psychostimulant drug that exerts persistent addictive effects. Most addictive drugs increase extracellular concentrations of dopamine (DA) in nucleus accumbens (NAc) and medial prefr.. Amphetamine addiction, conserved biosystem (from KEGG). Amphetamine addiction, conserved biosystem. Amphetamine is a psychostimulant drug that exerts persistent addictive effects. Most addictive drugs increase extracellular concentrations of dopamine (DA) in nucleus accumbens (NAc) and medial prefr.. Androgen receptor signaling pathway, organism- specific biosystem (from Wiki. Pathways). Androgen receptor signaling pathway, organism- specific biosystem. Androgens, mainly testosterone and 5alpha- dihydrotestosterone (DHT) play significant role in the growth and development of the male reproductive organs. These steroid hormones bring about their biolo.. Apoptosis, organism- specific biosystem (from Wiki. Pathways). Apoptosis, organism- specific biosystem. Apoptosis is a distinct form of cell death that is functionally and morphologically different from necrosis. Nuclear chromatin condensation, cytoplasmic shrinking, dilated endoplasmic reticulum, and .. Apoptosis Modulation and Signaling, organism- specific biosystem (from Wiki. Pathways). Apoptosis Modulation and Signaling, organism- specific biosystem. Apoptosis, or cell death program, can be activated by various mechanisms within the extrinsic and the intrinsic pathway. While activation of cell death receptors leads to the engagement of the extrin.. B Cell Receptor Signaling Pathway, organism- specific biosystem (from Wiki. Pathways). B Cell Receptor Signaling Pathway, organism- specific biosystem. The B cell receptor includes membrane ?? Ig alpha (CD7. 9A)/Ig beta (CD7. B) heterodimer. Activation of the B cell receptor involves phosphorylation o.. B cell receptor signaling pathway, organism- specific biosystem (from KEGG). B cell receptor signaling pathway, organism- specific biosystem. B cells are an important component of adaptive immunity. They produce and secrete millions of different antibody molecules, each of which recognizes a different (foreign) antigen. The B cell receptor.. B cell receptor signaling pathway, conserved biosystem (from KEGG). B cell receptor signaling pathway, conserved biosystem. B cells are an important component of adaptive immunity. They produce and secrete millions of different antibody molecules, each of which recognizes a different (foreign) antigen. The B cell receptor.. BCR signaling pathway, organism- specific biosystem (from Pathway Interaction Database). BCR signaling pathway, organism- specific biosystem. BCR signaling pathway. BDNF signaling pathway, organism- specific biosystem (from Wiki. Pathways). BDNF signaling pathway, organism- specific biosystem. Brain- derived neurotrophic factor (BDNF) is a neurotrophin essential for growth, differentiation, plasticity, and survival of neurons. BDNF is also required for processes such as energy metabolism, b.. CD4. 0/CD4. 0L signaling, organism- specific biosystem (from Pathway Interaction Database). CD4. 0/CD4. 0L signaling, organism- specific biosystem. CD4. 0/CD4. 0L signaling. CDC4. 2 signaling events, organism- specific biosystem (from Pathway Interaction Database). CDC4. 2 signaling events, organism- specific biosystem. CDC4. 2 signaling events. Calcineurin- regulated NFAT- dependent transcription in lymphocytes, organism- specific biosystem (from Pathway Interaction Database). Calcineurin- regulated NFAT- dependent transcription in lymphocytes, organism- specific biosystem. Calcineurin- regulated NFAT- dependent transcription in lymphocytes. Calcium signaling in the CD4+ TCR pathway, organism- specific biosystem (from Pathway Interaction Database). Calcium signaling in the CD4+ TCR pathway, organism- specific biosystem. Calcium signaling in the CD4+ TCR pathway. Cellular Senescence, organism- specific biosystem (from REACTOME). Cellular Senescence, organism- specific biosystem. Cellular senescence involves irreversible growth arrest accompanied by phenotypic changes such as enlarged morphology, reorganization of chromatin through formation of senescence- associated heterochr.. Cellular responses to stress, organism- specific biosystem (from REACTOME). Cellular responses to stress, organism- specific biosystem. Cells are subject to external molecular and physical stresses such as foreign molecules that perturb metabolic or signaling processes, and changes in temperature or p. H. The ability of cells and tissu.. Chagas disease (American trypanosomiasis), organism- specific biosystem (from KEGG). Chagas disease (American trypanosomiasis), organism- specific biosystem. Trypanosoma cruzi is an intracellular protozoan parasite that causes Chagas disease. The parasite life cycle involves hematophagous reduviid bugs as vectors. Once parasites enter the host body, they .. Chagas disease (American trypanosomiasis), conserved biosystem (from KEGG). Chagas disease (American trypanosomiasis), conserved biosystem. Trypanosoma cruzi is an intracellular protozoan parasite that causes Chagas disease. The parasite life cycle involves hematophagous reduviid bugs as vectors. Once parasites enter the host body, they .. Choline metabolism in cancer, organism- specific biosystem (from KEGG). Choline metabolism in cancer, organism- specific biosystem. Abnormal choline metabolism is emerging as a metabolic hallmark that is associated with oncogenesis and tumour progression. Following transformation, oncogenic signalling via pathways such as the RAS.. Choline metabolism in cancer, conserved biosystem (from KEGG). Choline metabolism in cancer, conserved biosystem. Abnormal choline metabolism is emerging as a metabolic hallmark that is associated with oncogenesis and tumour progression. Following transformation, oncogenic signalling via pathways such as the RAS.. Cocaine addiction, organism- specific biosystem (from KEGG). Cocaine addiction, organism- specific biosystem. Drug addiction is a chronic, relapsing disorder in which compulsive drug- seeking and drug- taking behavior persists despite serious negative consequences. There is strong evidence that the dopaminergic.. Cocaine addiction, conserved biosystem (from KEGG). Cocaine addiction, conserved biosystem. Drug addiction is a chronic, relapsing disorder in which compulsive drug- seeking and drug- taking behavior persists despite serious negative consequences. There is strong evidence that the dopaminergic.. Colorectal cancer, organism- specific biosystem (from KEGG). Colorectal cancer, organism- specific biosystem. Colorectal cancer (CRC) is the second largest cause of cancer- related deaths in Western countries. CRC arises from the colorectal epithelium as a result of the accumulation of genetic alterations in .. Colorectal cancer, conserved biosystem (from KEGG). Colorectal cancer, conserved biosystem. Colorectal cancer (CRC) is the second largest cause of cancer- related deaths in Western countries. CRC arises from the colorectal epithelium as a result of the accumulation of genetic alterations in .. ![]() Find Easter crafts to make with kids for the Easter holiday including an Easter basket, Easter eggs, Easter bonnet and other Easter activities for kids. Fun Easter Activities for Teachers - Easter Unit of Study Coloring pages, crafts for kids, puzzles, word searches, stories, book reviews, party games. Discover dozens of kids' Easter activities & ideas for a fun family Easter. Make it great holiday with egg-cellent crafts, games, recipes & egg decorating. Easter. Easter is an important Christian festival and holiday celebrating the resurrection of Jesus Christ. There are many different customs for celebrating Easter around the world, but the Friday before Easter Sunday (Good Friday) and the Monday after are given as a bank holiday in the UK. Early celebrations included the painting of eggs, which represented Jesus' empty tomb. Now we give Easter eggs - often chocolate - to friends and family, and have Easter egg hunts for eggs hidden by the Easter bunny. Activity Village has a huge selection of Easter activities for you to enjoy with your kids, including colouring pages, printables, jokes, crafts and puzzles! Explore below. Easter is not a fixed holiday and the date varies between 2. March and 2. 5th April. Easter Sunday will next be celebrated on 2. ![]() ![]() Easter is an important Christian festival and holiday celebrating the resurrection of Jesus Christ. There are many different customs for celebrating Easter around the world, but the Friday before Easter Sunday (Good Friday. Explore PediaStaff's board 'Easter Activities' on Pinterest, a visual bookmarking tool that helps you discover and save creative ideas | See more about Easter Eggs, Therapy and Easter. Easter Activities. Everything you need for a great Easter celebration with your kids: Easter crafts. Links to Other Easter Activity Websites. ![]() Easter and Spring Crafts and Activities for kids. Kindergarten, preschool, and elementary school crafts. Make wonderful, simple crafts with things found around the house. Easter Activity Pages![]() Complement Activation In The Parkinson S Disease Substantia Nigra An Immunohistochemical Study1/22/2017 Microglial inflammation in the parkinsonian substantia nigra: relationship to alpha- synuclein deposition. Abstract. Background. The role of both microglial activation and alpha- synuclein deposition in Parkinson's disease remain unclear. We have tested the hypothesis that if microglia play a primary role in Parkinson's disease pathogenesis, the microglial "activated" phenotype should be associated with histopathological and/or clinical features of the disease. Methods. We have examined microglial MHC class II expression, a widely used marker of microglial activation, the occurrence of CD6. Parkinson's disease (PD). Using semi- quantitative severity ratings, we have examined the relationship between microglial activation, alpha- synuclein deposition, classical neuropathological criteria for PD, subtype of the disease and clinical course. Results. While we did not observe an association between microglial MHC class II expression and clinical parameters, we did find a correlation between disease duration and the macrophage marker CD6. In addition, we observed a significant correlation between the degree of MHC class II expression and alpha- synuclein deposition in the substantia nigra in PD. Conclusion. While microglia appeared to respond to alpha- synuclein deposition, MHC class II antigen expression by microglia in the substantia nigra cannot be used as an indicator of clinical PD severity or disease progression. . (MPTP). Immunohistochemical staining of dopaminergic neurons in. there is still a possibility that MPTP-induced complement activation does not. Complement activation in the Parkinson's disease substantia nigra.![]() ![]() In addition, a contributory or even causative role for microglia in the neuronal loss associated with PD as suggested by some authors seems unlikely. Our data further suggest that an assessment of microglial activation in the aged brain on the basis of immunohistochemistry for MHC class II antigens alone should be done with caution. Introduction. Parkinson's Disease (PD) is a common neurodegenerative disorder with the cardinal clinical features of tremor, rigidity, bradykinesia and loss of postural reflexes. Neuropathologically, the disease is characterized by a marked loss of dopaminergic neurons in the substantia nigra pars compacta (SN) and the presence of alpha- synuclein (a. SN)- positive Lewy bodies (LBs) in neurons of this and other brain areas also affected by nerve cell death. Plaque Complement Activation and Cognitive Loss in Alzheimer's Disease. Complement activation in the Parkinson's disease substantia nigra: an immunocytochemical study. J Neuroinflammation 2006. Relationship between microglial activation and dopaminergic neuronal loss in the substantia nigra: a time course study in a. Parkinson’s disease. SNc. substantia nigra. neuronal loss and activation of. An international consensus definition of Lewy body diseases on the basis of molecular as well as morphological criteria, which takes into account a. SN status of the brain, has been published recently http: //www. ICDNS. org[1]. The discovery of a. SN mutations and gene amplification in some familial forms of PD [2- 6]] and the identification of this protein as a major component of Lewy bodies (LBs) in common sporadic PD [7], has spurred interest in the role of a. SN in the pathophysiology of PD and other synucleinopathies. However, no direct causal relationship has yet been established between a. SN aggregation and the selective neuronal cell death characteristic of PD. LBs are also found incidentally in aged brain in the absence of other pathological features and without a clinical history of parkinsonism or dementia [8]. Attempts have been made to link the clinical progression of PD to the presence of a. SN inclusions and an anatomical staging model has been proposed [9], but the latter has been questioned by subsequent studies in which clinical data were also taken into account [1. ![]()
Apart from well established morphological criteria, activated microglia can be defined in tissue sections on the basis of the expression of several immune function- related proteins, notably complement receptors and MHC class II antigens (MHCII). Phagocytic activity and cytotoxic properties are usually considered end stages of microglial activation, at which point they are phenotypically indistinguishable from blood- borne macrophages. Activated microglia are associated with a large range of neurological insults from trauma and infection to autoimmune conditions, and their presence represents a common finding also in neurodegenerative disorders [1. ![]() ![]() However there is little knowledge about the molecular processes that mediate microglial activation and exactly which biological consequences may result from their enhanced state of "immune alertness" within affected CNS tissue. A transcriptome signature of interferon- gamma activated microglia has been provided recently [1. Microglial phenotypic changes have also been observed in normal aged individuals [1. Thus, "microglial senescence" confounds the problem of a definition of microglial activation in disease states, and in neurodegenerative diseases in particular which are often age- related, as no specific causative stimulus has been identified in the process. While microglia clearly show changes in their phenotypic profile in neurodegeneration, it is by no means clear whether they are actively involved in the progression of PD. Microglia- derived macrophages can be found in the PD SN, and neuromelanin pigment taken up from degenerated dopaminergic nerve cells is characteristically observed in SN phagocytes. In animal models of nigrostriatal degeneration using 6- hydroxydopamine and 1- methyl- 4- phenyl- 1,2,3,6- tetrahydropyridine (MPTP), inhibition or attenuation of the microglial immune response increases neuronal survival. However, those results have so far not been replicated in clinico- pathological studies, and the simple chemical lesions currently employed in animal studies by all likelihood do not fully reflect the chronic neurodegenerative disease process in humans [1. In the present study, we independently evaluate the severity of alpha- synuclein deposition and microglial activation identified by immunohistochemical staining in the SN in a large cohort of clinically and pathologically confirmed PD cases. We have studied the microglial response in PD on two levels, by observing MHCII - immunoreactive cells (putatively activated microglia but possibly only senescent cells) and CD6. Materials and methods. Parkinson's disease cases. PD nigrae were evaluated immunohistochemically. UK Parkinson's Disease Society Tissue Bank at Imperial College London (PDSTB). Additional tissue sections from 1. Institute of Neuropathology, University of Munich, Germany. These Parkinson's cases had been previously diagnosed, neuropathologically screened for confounding pathology, and examined in a study of apoptosis and microglial activation [1. Archival sections were immunolabelled for alpha- synuclein (see below) and used as a control group to ensure that variation within our PDSTB cohort was within an established range. Clinical and neuropathological assessment of cases. For the PDSTB cohort, clinical reports were evaluated in detail by an experienced neurologist with a special interest in Parkinson's disease (RKBP). Neuropathological assessment was based on slides provided by the PDSTB for alpha- synuclein, tau and beta- amyloid immunohistochemistry of superior frontal gyrus, the hippocampal region and midbrain as minimum data sets, and screening of the cases for confounding pathology was based on hematoxylin and eosin examination of a standard series of 1. Nine cases showed varying degrees of concurrent Alzheimer's disease (AD)- type pathology (tau- immunopositive tangles and/or beta- amyloid- immunopositive plaques) of isocortical and/or entorhinal type ranging from grades 1–3 http: //www. ICDNS. org. Three cases were excluded based on a final neuropathological diagnosis of AD, progressive supranuclear palsy (PSP), and young- onset familial PD, respectively, leaving a cohort of 1. Munich and PDSTB groups (Table 1). Immunohistochemical evaluation of protein levels. Immunohistochemical reactions were performed using the avidin- biotin complex (ABC)/peroxidase method with mouse monoclonal antibodies anti- human HLA- DP, DQ, DR (clone CR3/4. Dako, dilution 1/1. Becton- Dickinson, dilution 1/3. For the PDSTB group, additional immunohistochemistry was carried out with anti- CD6. PGM1, Dako, dilution 1/2. Sections were dewaxed in xylene, rehydrated, and endogenous peroxidase activity was blocked by 3. Antigen unmasking consisted of boiling in 0. M EDTA (2. 0 min. W in microwave) and 1. HLA- DP, DQ, DR and anti- alpha- synuclein, respectively. No antigen unmasking was used with anti- CD6. Slides were then incubated in primary antibody diluted in phosphate- buffered saline (PBS) overnight at 4°C. The following day, after washing in PBS, they were incubated in horse- anti- mouse secondary antibody (Vector, dilution 1/2. ABC complex (Vector, dilution 1/2. Immunoreactivity was visualised with 3,3'- diaminobenzidine (Vector kit). After immunohistochemical staining, sections were given semi- quantitative severity ratings for a. SN, MHCII, and CD6. EC and MBG) blinded to case number. The SN was defined as the area extending laterally from the exit of the third nerve, superior to the cerebral peduncle and inferior to the medial lemniscus, ideally at the height of the red nucleus with the presence of melanised neurons or their remnants indicating the main region of interest. The severity ratings were determined across the entire region of SN, based on the density of immunopositive structures, with 0 (none), 1 (mild), 2 (moderate) and 3 (high). For a. SN, both intra- and extra- cellular inclusions were considered provided they fell within the immediate area of the substantia nigra. This was particularly relevant in areas of severe neuronal loss, often encountered more laterally, where significant alpha- synuclein pathology could still be observed. The morphological variation in a. SN deposition was not assessed, simply the frequency of events. All clearly identifiable a. SN- immunoreactive structures, including LBs, neurites, fibrils, and smaller, punctate formations, were considered. For microglial response, severity was judged primarily by immunoreactivity, however morphology was taken into account in that perivascular immunoreactivity was excluded. Fun and Creative Activities For Kids. Check-out our growing list of kids' activities and find something. Many children begin writing or 'play writing' well. At Creative Activities For Kids you will find endless ways to replace your. It is good to have an indoor activities list. Creative summer activities for. List of Christmas Activities. allowing children to explore their creativity and. inexpensive and creative way to bring the colours of Christmas and fun into. Your Very Own List of Creative Activities 1. Read children's stories and record. List of Possible Sites for Random Photo night. Fun and Creative Activities For Kids. Kindergarten Testing Explained. There was a time not that long ago that when a child turned a certain age, they simply went to kindergarten, whether they had any pre- school behind them or not. Now you may find that they need to take a kindergarten test. (continue reading)Three Early Childhood Writing Activities. Many children begin writing or "play writing" well before they can read even the simplest words. Here are three early childhood writing activities you can use to guide your child's writing enthusiasm and help develop their creative talents. (continue reading)The Most Important Factor For School Readiness. According to Northwestern University researcher Greg Duncan, "We find the single most important factor in predicting later academic achievement is that children begin school with a mastery of early math and literacy concepts." (continue reading)Assessing Early Literacy Skills. Think your child may be ready to start reading? A child with a head start on reading and literacy has a greater chance of success in the first years of school and is more likely to finish college compared to children who begin to read at a later age. (continue reading)Expanding Vocabulary with Rhyming Games. Rhyming is one of the first skills that children can practice to start understanding letter- sound- word relationships and can be a fun way to expand early vocabulary by building on familiar word patterns that children are already comfortable with. (continue reading)Reading To Your Child - 1. Reasons To Do It. Creative activities: visual art and construction. You don’t always need to give your child new play materials. Using everyday objects, and making it up as you go.Few dispute the benefits of reading aloud to young children, and there are many reasons that reading to children is important. Here are 1. Explore ways to encourage creativity in kids and further creative play. At PBS Parents we let creativity blossom! Children & Media. Getting creative doesn't need to cost a. The children can take it in turns to play DJ and select different. There are stacks of activities they could do. Creative Classroom Activities is the second of a two-volume anthology of articles published between 1989. Add to My Resource List. Remove from My Resource List. Many of these boot disk utilities are familiar Windows functions: Partition Manager (DiskPart.exe), Check Disk, Command Prompt. Active@ Password Changer Active@ Boot Disk(Live CD) Active@ Partition Manager Active@ Hard Disk Monitor NTFS Reader for DOS Active@ Kill Disk Active@ ERASER. This is Active@ Boot Disk version history page. Active@ Boot Disk is a complete and functioning computer operating system on CD/DVD/USB disk. Active@ Boot Disk contains a collection of powerful utilities to perform data recovery, data imaging, secure data erasure and a number of other tasks all from a. Active@ Partition Recovery. Active@ Partition Recovery is a data recovery software tool that helps to recover deleted partitions and logical disks on hard drives. Active@ Boot Disk is a bootable CD that gives you a lightweight Windows 7 (WinPE 3.0) environment with disk image, data recovery, password resetting, data erasure and. Active@ Boot Disk. Active@ Boot Disk is a complete and functioning computer operating system on CD/DVD/USB disk. Active@ Boot Disk does not modify the operating. Boot Disk System Utilities. One of the main differences between the Active@ Boot Disk utilities and Windows utilities is that with Active@ Boot Disk you can access them even if your hard drive is damaged. Displays system information about your local machine. An applet that helps you create and display a status report for your hard drive. Choose the resolution and number of colors displayed. Get information about computer performance and details about running applications, processes, network activity, user and system services. View or modify currently running services. View the full name of a process. Monitor applications and services running system performance and resources. Change your computer’s date, time and time zone. Search for and display specified registry details or create a custom registry hive. After you find the registry hive that you are looking for, you can edit it with Registry Editor. Add language support for a second language. This includes special keyboard layout and the ability to display language characters on the screen. You can use a different language for a file name or when recording notes about changes made to the system. View and edit entries in BCD (Boot Configuration Data) store to make a Windows system partition bootable Active@ Boot Disk System Utilities. Alfresco integration with Active Directory. One of the main features of the Alfresco ECM System is the ability to integrate user authentication and synchronization with Microsoft Active Directory. Unfortunately, integration is not trivial and it is error prone. While I’ll provide explanations how stuff works you can also have a look at the chapter 6 of the Professional Alfresco: Practical Solutions for Enterprise Content Management (Wrox Programmer to Programmer) [amazon. Alfresco 3 Enterprise Content Management Implementation [amazon. In this guide I’ll show you how to achieve full integration with Active Directory which includes Alfresco Explorer and Alfresco Share SSO, CIFS SSO, and Active Directory (LDAP) users and groups synchronization. Prerequisites for this article are: working Active Directory environment, functional basic installation of Alfresco i. Alfresco using the default admin/admin user. ![]() I tend to use bundled Alfresco/Tomcat zip installation since installer can be buggy from version to version and you don’t have full control. I recommend using Alfresco CE 3. NTLM, CIFS and LDAP merged from enterprise version. In my case Alfresco is installed on Windows 2. EE with SP2 3. 2bit. Alfresco subsystems. Introduced in version 3. Alfresco subsystems: Authentication. Synchronization. File Servers. IMAPsys. Admin. WCM Deployment Receiver. Third party (Open. Office, Image Magick, etc.)Subsystems in their nature are separate configurable modules responsible for a sub- part of the Alfresco functionality. Subsystems that we are interested in this article are the first three: authentication, synchronization and file servers. ![]() Note that configuration for subsystems is located in their respective folders as shown in the picture: Note that LDAP synchronization is linked to LDAP authentication. Lets get our hands dirty now. Authentication. First thing to do to integrate user authentication with Active Directory is to configure the main authentication chain in Alfresco configuration. Join CentOS/RedHat to Active Directory Domain Sometimes you have to join Linux to an Active Directory Domain, If you run a Full UNIX/Linux environment, a better. Synchronization. For user synchronization to work we must configure Alfresco to hit the Active Directory domain controller with the appropriate LDAP queries. ![]() We do this by adding the following line in \Alfresco\tomcat\shared\classes\alfresco- global. Note: ldap- ad is here in chain to support later explained user synchronization. Now, create the file \subsystems\Authentication\passthru\passthru. Local. Server=falsepassthru. DOMAIN\\1. 92. 1. Guest. Login=falsentlm. Unknown. User. To. Guest=falsepassthru. CIFS=truepassthru. FTP=falsepassthru. Access=falsepassthru. Administrator. User. Names=AD_usernames. What we tell Alfresco with the above config is to use the external server for authentication, enable NTLM SSO, and put the list of usernames that should have the administrator role. Now, edit the \Alfresco\tomcat\shared\classes\alfresco\web- extension\webscript- framework- config- custom. Remote”> < remote> < !– SSL client certificate + trusted CAs. Optionally used to authenticate share to an external SSO system such as CAS –> < keystore> < path> alfresco/web- extension/alfresco- system.
Cookie< /id> < name> Alfresco Connector< /name> < description> Connects to an Alfresco instance using cookie- based authentication< /description> < class> org. Alfresco. Connector< /class> < /connector> < endpoint> < id> alfresco< /id> < name> Alfresco – user access< /name> < description> Access to Alfresco Repository Web. Scripts that require user authentication< /description> < connector- id> alfresco. Cookie< /connector- id> < endpoint- url> http: //localhost: 8. Next, edit the WEB- INF\web. Authentication Filter< /filter- name> < filter- class> org. NTLMAuthentication. Filter< /filter- class> < init- param> < param- name> endpoint< /param- name> < param- value> alfresco< /param- value> < /init- param> < /filter> < filter- mapping> < filter- name> Authentication Filter< /filter- name> < url- pattern> /page/*< /url- pattern> < /filter- mapping> < filter- mapping> < filter- name> Authentication Filter< /filter- name> < url- pattern> /p/*< /url- pattern> < /filter- mapping> < filter- mapping> < filter- name> Authentication Filter< /filter- name> < url- pattern> /s/*< /url- pattern> < /filter- mapping> SSO should now be fully functional for Alfresco Explorer and Alfresco Share. Lets synchronize the users and groups now. Synchronization. For user synchronization to work we must configure Alfresco to hit the Active Directory domain controller with the appropriate LDAP queries. Queries are highly dependant of ones AD structure so the following configuration covers the scenario where: All groups that I want in Alfresco are members of a single group called Alfresco Groups,Users that I want synchronized to Alfresco are members of the above mentioned member groups. This structure is nice if you have users in different OUs but don’t want them all to be also present in Alfresco. So, edit the \subsystems\Authentication\ldap- ad\ldap- ad. LDAP Sync## This flag enables use of this LDAP subsystem for authentication. It may be# that this subsytem should only be used for synchronization, in which case# this flag should be set to false. This flag enables use of this LDAP subsystem for user and group# synchronization. It may be that this subsytem should only be used for# authentication, in which case this flag should be set to false. Name. Format=%sldap. Guest. Login=trueldap. Is there any way to mount a remote CIFS/SMB/SAMBA share as a folder/directory and not as a drive letter. For example, I want this map: \\Server\ShareName -> C:\Folder. ![]() The default principal to bind with (only used for LDAP sync). This should be a UPN or DNldap. The password for the default principal (only used for LDAP sync)ldap. Your. Pass# If positive, this property indicates that RFC 2. This# overcomes any size limits imposed by the LDAP server. Batch. Size=1. 00. The query to select all objects that represent the groups to import. ![]() Query=(& (objectclass\=group)(member. Of\=cn\=Alfresco Groups,ou\=user,dc\=domain,dc\=local))# The query to select objects that represent the groups to import that have changed since a certain time. Differential. Query=(& (objectclass\=group)(member. Of\=cn\=Alfresco Groups,ou\=user,dc\=domain,dc\=local)(!(modify. Timestamp< \={0})))# The query to select all objects that represent the users to import. Query=(& (objectclass\=user)(|(member. Of\=CN\=Developers,OU\=user,DC\=domain,DC\=local)(member. Of\=CN\=Sales,OU\=user,DC\=domain,DC\=local))(user. Account. Control\: 1. The query to select objects that represent the users to import that have changed since a certain time. Differential. Query=(& (objectclass\=user)(|(member. Of\=CN\=Developers,OU\=user,DC\=domain,DC\=local)(member. Of\=CN\=Sales,OU\=user,DC\=domain,DC\=local))(user. Account. Control\: 1. Timestamp< \={0})))# The group search base restricts the LDAP group query to a sub section of tree on the LDAP server. Search. Base=dc\=domain,dc\=local# The user search base restricts the LDAP user query to a sub section of tree on the LDAP server. Search. Base=dc\=domain,dc\=local# The name of the operational attribute recording the last update time for a group or user. Timestamp. Attribute. Name=modify. Timestamp# The timestamp format. Unfortunately, this varies between directory servers. Format=yyyy. MMdd. HHmmss’. 0. Z’# The attribute name on people objects found in LDAP to use as the uid in Alfrescoldap. Id. Attribute. Name=s. AMAccount. Name# The attribute on person objects in LDAP to map to the first name property in Alfrescoldap. First. Name. Attribute. Name=given. Name# The attribute on person objects in LDAP to map to the last name property in Alfrescoldap. Last. Name. Attribute. Name=sn# The attribute on person objects in LDAP to map to the email property in Alfrescoldap. Email. Attribute. Name=mail# The attribute on person objects in LDAP to map to the organizational id property in Alfrescoldap. Organizational. Id. Attribute. Name=company# The default home folder provider to use for people created via LDAP importldap. Home. Folder. Provider=user. Homes. Home. Folder. Provider# The attribute on LDAP group objects to map to the gid property in Alfrecsoldap. Id. Attribute. Name=cn# The group type in LDAPldap. Type=group# The person type in LDAPldap. Type=user# The attribute in LDAP on group objects that defines the DN for its membersldap. Member. Attribute. Name=membersynchronization. Changes. Only=true. What we do above is: disable LDAP authentication and enable just LDAP synchronization, set the LDAP URL to query, credentials with read rights on LDAP, groups query, users query, and LDAP attributes to map Alfresco user’s attributes to. Note that LDAP synchronization is not instant, duration depends on number of objects to synchronize and is scheduled to run every hour (configurable in scheduled- jobs- context. CIFSNow, the problematic part. CIFS is usually the hardest one to get it to work properly. So lets start with basic config that should work and then try to fix the eventual problems if there are any. Edit the \subsystems\file. Servers\default\default\changes. Alfrescofilesystem. Mappings=DOMAINfilesystem. Mappings. value. DOMAIN. From=1. 92. 1. 68. Mappings. value. DOMAIN. To=1. 92. 1. 68. 0. HOSTNAMEcifs. domain=DOMAINcifs. If it doesn’t work you can try to enable only the Java socket based Net. BIOS over TCP/IP. You do this by copying file- servers- context. Servers\default\default\custom- file- servers- context. BIOSSMB”> < bean class=”org. Net. BIOSSMBConfig. Bean”> < property name=”bind. To”> < value> ${cifs. Port”> < value> ${cifs. BIOSSMB. session. Port}< /value> < /property> < property name=”name. Port”> < value> ${cifs. BIOSSMB. name. Port}< /value> < /property> < property name=”datagram. Port”> < value> ${cifs. BIOSSMB. datagram. Port}< /value> < /property> < property name=”platforms”> < value> linux,solaris,macosx< /value> < /property> < /bean> < /property> –> < !– Use Win. ![]() Conversion of factor VII to factor VIIa is catalyzed. [7] Factor VII activity is influenced by mutations of. et al. Effect of recombinant activated factor VII on perioperative blood loss in patients. ![]() ![]() Medscape: Medscape Access. This Privacy Policy applies to the websites www. 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Factor VII (EC 3.4.21.21, blood-coagulation factor VIIa, activated blood coagulation factor VII, formerly known as proconvertin) is one of the proteins that causes blood to clot in the coagulation cascade. It is an enzyme of. The use of recombinant activated factor VII in platelet disorders: A critical review of the literature on ResearchGate, the professional network for scientists. Blood Transfus 2009; 7: 24-8 DOI 10.2450/2008.0015-08. Original Article from The New England Journal of Medicine — Recombinant Activated Factor VII for Acute Intracerebral Hemorrhage. Recombinant activated factor VII Drug Information from Drugs.com. Includes Recombinant activated factor VII side effects, interactions and indications. 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MD and market research surveys conducted on behalf of third party sponsors either by Web. MD or a third party ("Sponsored Surveys"), where in this context "sponsor" means the provider of products and services who commissioned the survey. Web. MD surveys may be administered by one of our companies or by a third party acting on our behalf. For some surveys, you may be asked to provide contact information for honoraria fulfillment. Continuing Medical Education. Activated Factor 7 Blood Disease![]() Traumatic bleeding with recombinant factor VIIa. Lancet 1999;354:1879. 7) Mayer SA, Brun NC, Begtrup K, Broderick. et al. Effect of recombinant activated factor VII on perioperative blood loss in patients undergoing retropubic. Recombinant activated factor VII in post partum haemorrhage. Thoughts on the mechanism of action of high-dose activated Factor VII. Blood. Popham P, Ogle R. Guidelines for the use of recombinant activated factor VII. 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How We May Use Your Information. ![]() The latest critic and user reviews, photos and cast info for Criminal Activities. SXSW Film Festival; Tribeca; Cannes; Comic-Con; Emmy Awards; Venice Film Festival. Review: Criminal Activities, Now On Blu-ray And VOD 16 February 2016 11:00 AM. The presence of John Travolta isn't the only element reminiscent of Quentin Tarantino in actor Jackie Earle Haley's directorial debut. Featuring self-consciously. Criminal Activities’ Review: Jackie Earle Haley’s Derivative Directing Debut. John Travolta as a charismatic gangster in an impeccably tailored suit. Casual references to Marcel Proust and quantum physics. An outrageously byzantine plot orchestrated around a big twist ending. The ’9. 0s are back again in “Criminal Activities,” a profane thriller that so closely resembles the B- movies that followed “The Usual Suspects,” “Pulp Fiction” and “Get Shorty,” it could be mistaken for an archeological discovery. Making his directorial debut, veteran thesp Jackie Earle Haley uncannily re- creates the feeling of watching an Elmore Leonard clone circa 1. There’s nothing remotely fresh about this revival, but tight pacing and an overqualified cast keep things zipping along nicely, which may not be enough to draw people to theaters on Nov. В 2. 0, but should clear the lower bar of its simultaneous VOD release. В Were the four buddies at the center of “Criminal Activities” not texting so furiously, the film could be mistaken for another “Suicide Kings” or “Things to Do in Denver When You’re Dead” or “Two Days in the Valley,” since so little has been invested in the contemporary backdrop. All the emphasis is on the foreground, which offers the familiar premise of amateur crooks getting mixed up with trigger- happy professionals. Playing the type of guy who wears shades indoors, Michael Pitt stars as Zach, a smug young stock broker who reconnects with his friends Noah (Dan Stevens), Warren (Christopher Abbott), and Bryce (Rob Brown) at a funeral. Review: A bit meh - If John. Criminal Activities had me glued to the screen for. wittiness without much success and you have Criminal Activities. This isn't a. Jackie Earle Haley's Criminal Activities is a throwback to the post-Tarantino imitations that flourished in the 1990s in the wake of Pulp Fiction's success. The '90s are back again in “Criminal Activities,” a profane thriller that so closely resembles the B-movies that followed “The Usual Suspects. A few drinks later, the boys start scheming. Noah proposes a can’t- miss insider trading scheme involving a small pharmaceutical company on the verge of FDA approval for a game- changing drug. The four agree to partner up on a $2. Noah has money on hand, which the others assume he inherited from his recently deceased father. When the company goes under, they discover the source of Noah’s cash is Eddie (John Travolta), a mob kingpin who sips on a kale shake and recites the seven rules of economics before informing them they owe him twice the money they borrowed. Eddie offers the cash- strapped men a way out: Kidnap a vicious gangster named Marques (Edi Gathegi), a member of a rival crime syndicate. ![]() Criminal Activities” aims for a mix of comic ineptitude and two- fisted gangster theatrics, and Haley hits the mark in the kidnapping sequence, which has Marques nearly muscling his way past the hapless quartet in a public bathroom. Stevens’ about- face from the masculine force of “The Guest” to a bushy- haired weakling here is a marvel to behold, but it’s Gathegi who steals the movie as a cool operator who’s completely in control of the situation, even when pinned to a chair with duct tape. Travolta’s return to Chili Palmer mode isn’t a bad idea, but the glib one- liners in Robert Lowell’s script don’t give him much of an arsenal; Gathegi’s serene posture and missile- guided barbs present the more clear and present danger in the room. Pitt and the gang have a much harder time of it. Each of the four have the thinnest of backstories (one frets that his girlfriend is cheating on him, another struggles with his sobriety, etc.), but once the trouble escalates, they mostly just scream at each other. Haley and Lowell give the peripheral characters a little Elmore Leonard color, but they aren’t as generous with Pitt, Stevens, Abbott, and Brown, who tell each other to “Shut the f—В up! Marques doesn’t want to spend another minute in the room with these guys; after a while, the feeling is mutual. Haley gives himself the type of role Jackie Earle Haley might play as Eddie’s chief goon. Tech credits are serviceable across the board, though the pic’s habit of goosing up the action with brief song clips reps a minor but persistent irritation. Reviewed online, Chicago, Nov. Running time: 9. 4 MIN. В Production. An RLJ Entertainment release of a Phoenix Rising and Wayne Rice production in association with May Day Movie Prods., Nee. Nee Prods., Producer Capital Fund, and Jackie Earle Haley. ![]() The first notion presented in Criminal Activities, the directorial debut of Watchmen's Jackie Earle Haley, is a common and precise one: life is. The '90s are back again in “Criminal Activities,” a profane thriller that so closely resembles the B-movies that followed “The Usual Suspects,” “Pulp. Produced by Wayne Rice, Howard Burd, Micah Sparks. Executive producers, Mark B. Johnson, Daniel Diamond, Roger May, Mario Coluccio, Gayle Nosal, Chris Haley, Michael Pitt, Dan Stevens, John Travolta, Michael Brickman, Blair Hahn, Warren Ostergard. Co- producers, Linda Favilia, Anson Downes. Co- executive producers, Gary Dankelfsen, Cherelle George. Crew. Directed by Jackie Earle Haley. Screenplay, Robert Lowell. Camera (color, widescreen, HD), Seamus Tierney; editor, Alex Marquez; music, Keefus Ciancia; music supervisor, Sean Mulligan; production designer, Jennifer Klide; set decorator, Carmen Navis; costume designer, Carla Shivener; sound (Dolby Digital), Marlowe Taylor; sound designer, Jeffrey Alan Pitts; re- recording mixer, Michael Perricone; visual effects supervisor, Chris Haley; stunt coordinator, Richard Fike Zach; assistant director, Chris De. Benedetto; casting, Barbara Mc. Carthy, Angela Demo. With. Michael Pitt, Dan Stevens, Christopher Abbott, Rob Brown, Edi Gathegi, Jackie Earle Haley, John Travolta. ![]() How to Calculate Specific Activities of Enzymes. Enzymatic activity is the rate at which an enzyme. The specific activity of an enzyme. Enzyme assay - Wikipedia, the free encyclopedia. Beckman DU6. 40 UV/Vis spectrophotometer. Enzyme assays are laboratory methods for measuring enzymatic activity. They are vital for the study of enzyme kinetics and enzyme inhibition. Enzyme units[edit]Amounts of enzymes can either be expressed as molar amounts, as with any other chemical, or measured in terms of activity, in enzyme units. Calculating specific activity of an enzyme? Calculating the total enzyme activity and specific activity.? How to calculate specific activity for enzymes? BISC220/S10: Sample Calculations Lab 2. From OpenWetWare. Jump to: navigation, search. Wellesley College BISC 220 Cellular. ONPG assay for enzyme specific activity. Calculation of Enzyme activities. for calculation of enzymes particularly specific activity of enzyme : change in OD/time taken (min). Enzyme activity[edit]Enzyme activity = moles of substrate converted per unit time = rate Г— reaction volume. Enzyme activity is a measure of the quantity of active enzyme present and is thus dependent on conditions, which should be specified. The SI unit is the katal, 1 katal = 1 mol sв€’1, but this is an excessively large unit. A more practical and commonly used value is enzyme unit (U) = 1 Ојmol minв€’1. Specific activity is the activity per quantity of a radionuclide and is a physical property of that radionuclide. [1] [2] Activity is a quantity related to. This Site Might Help You. RE: Calculating specific activity of enzyme? I have been given the following info but the protein volume and the end units of SA. As i said in my previous post, specific activity is defined as units of activity per unit of enzyme. in your first post you say that you already have this value. Specific activity calculations. Enzyme Assay, Enzyme Activity and Specific Activity - Duration. Quick Guide to Calculating Enzyme Activity. If the specific activity of 100% pure enzyme is known, then an impure sample will have a lower specific activity, allowing purity to be calculated. Types of assay All. U corresponds to 1. Enzyme activity as given in katal generally refers to that of the assumed natural target substrate of the enzyme. Enzyme activity can also be given as that of certain standardized substrates, such as gelatin, then measured in gelatin digesting units (GDU), or milk proteins, then measured in milk clotting units (MCU). The units GDU and MCU are based on how fast one gram of the enzyme will digest gelatin or milk proteins, respectively. GDU equals approximately 1. MCU.[2]An increased amount of substrate will increase the rate of reaction with enzymes, however once past a certain point, the rate of reaction will level out because the amount of active sites available has stayed constant. Specific activity[edit]The specific activity of an enzyme is another common unit. This is the activity of an enzyme per milligram of total protein (expressed in Ојmol minв€’1mgв€’1). Specific activity gives a measurement of enzyme purity in the mixture. It is the moles of product formed by an enzyme in a given amount of time (minutes) under given conditions per milligram of total proteins. Specific activity is equal to the rate of reaction multiplied by the volume of reaction divided by the mass of total protein. The SI unit is katal kgв€’1, but a more practical unit is Ојmol mgв€’1 minв€’1. Specific activity is a measure of enzyme processivity, at a specific (usually saturating) substrate concentration, and is usually constant for a pure enzyme. For elimination of errors arising from differences in cultivation batches and/or misfolded enzyme etc. This is a measure of the amount of active enzyme, calculated by e. The specific activity should then be expressed as Ојmol minв€’1 mgв€’1 active enzyme. If the molecular weight of the enzyme is known, the turnover number, or Ојmol product per second per Ојmol of active enzyme, can be calculated from the specific activity. The turnover number can be visualized as the number of times each enzyme molecule carries out its catalytic cycle per second. Related terminology[edit]The rate of a reaction is the concentration of substrate disappearing (or product produced) per unit time (mol Lв€’1 sв€’1). The % purity is 1. Г— (specific activity of enzyme sample / specific activity of pure enzyme). The impure sample has lower specific activity because some of the mass is not actually enzyme. If the specific activity of 1. Types of assay[edit]All enzyme assays measure either the consumption of substrate or production of product over time. A large number of different methods of measuring the concentrations of substrates and products exist and many enzymes can be assayed in several different ways. Biochemists usually study enzyme- catalysed reactions using four types of experiments: [3]Initial rate experiments. When an enzyme is mixed with a large excess of the substrate, the enzyme- substrate intermediate builds up in a fast initial transient. Then the reaction achieves a steady- state kinetics in which enzyme substrate intermediates remains approximately constant over time and the reaction rate changes relatively slowly. Rates are measured for a short period after the attainment of the quasi- steady state, typically by monitoring the accumulation of product with time. Because the measurements are carried out for a very short period and because of the large excess of substrate, the approximation that the amount of free substrate is approximately equal to the amount of the initial substrate can be made. The initial rate experiment is the simplest to perform and analyze, being relatively free from complications such as back- reaction and enzyme degradation. It is therefore by far the most commonly used type of experiment in enzyme kinetics. Progress curve experiments. In these experiments, the kinetic parameters are determined from expressions for the species concentrations as a function of time. The concentration of the substrate or product is recorded in time after the initial fast transient and for a sufficiently long period to allow the reaction to approach equilibrium. Progress curve experiments were widely used in the early period of enzyme kinetics, but are less common now. Transient kinetics experiments. In these experiments, reaction behaviour is tracked during the initial fast transient as the intermediate reaches the steady- state kinetics period. These experiments are more difficult to perform than either of the above two classes because they require specialist techniques (such as flash photolysis of caged compounds) or rapid mixing (such as stopped- flow, quenched flow or continuous flow). Relaxation experiments. In these experiments, an equilibrium mixture of enzyme, substrate and product is perturbed, for instance by a temperature, pressure or p. H jump, and the return to equilibrium is monitored. The analysis of these experiments requires consideration of the fully reversible reaction. Moreover, relaxation experiments are relatively insensitive to mechanistic details and are thus not typically used for mechanism identification, although they can be under appropriate conditions. Enzyme assays can be split into two groups according to their sampling method: continuous assays, where the assay gives a continuous reading of activity, and discontinuous assays, where samples are taken, the reaction stopped and then the concentration of substrates/products determined. Temperature- controlled cuvette holder in a spectrophotometer. Continuous assays[edit]Continuous assays are most convenient, with one assay giving the rate of reaction with no further work necessary. There are many different types of continuous assays. Spectrophotometric[edit]In spectrophotometric assays, you follow the course of the reaction by measuring a change in how much light the assay solution absorbs. If this light is in the visible region you can actually see a change in the color of the assay, and these are called colorimetric assays. The MTT assay, a redox assay using a tetrazolium dye as substrate is an example of a colorimetric assay. UV light is often used, since the common coenzymes NADH and NADPH absorb UV light in their reduced forms, but do not in their oxidized forms. An oxidoreductase using NADH as a substrate could therefore be assayed by following the decrease in UV absorbance at a wavelength of 3. Direct versus coupled assays. Even when the enzyme reaction does not result in a change in the absorbance of light, it can still be possible to use a spectrophotometric assay for the enzyme by using a coupled assay. Here, the product of one reaction is used as the substrate of another, easily detectable reaction. For example, figure 1 shows the coupled assay for the enzyme hexokinase, which can be assayed by coupling its production of glucose- 6- phosphate to NADPH production, using glucose- 6- phosphate dehydrogenase. Fluorometric[edit]Fluorescence is when a molecule emits light of one wavelength after absorbing light of a different wavelength. Fluorometric assays use a difference in the fluorescence of substrate from product to measure the enzyme reaction. These assays are in general much more sensitive than spectrophotometric assays, but can suffer from interference caused by impurities and the instability of many fluorescent compounds when exposed to light. An example of these assays is again the use of the nucleotide coenzymes NADH and NADPH. Here, the reduced forms are fluorescent and the oxidised forms non- fluorescent. Oxidation reactions can therefore be followed by a decrease in fluorescence and reduction reactions by an increase.[5] Synthetic substrates that release a fluorescent dye in an enzyme- catalyzed reaction are also available, such as 4- methylumbelliferyl- ОІ- D- galactoside for assaying ОІ- galactosidase. Calorimetric[edit]. Chemiluminescence of Luminol. Calorimetry is the measurement of the heat released or absorbed by chemical reactions. These assays are very general, since many reactions involve some change in heat and with use of a microcalorimeter, not much enzyme or substrate is required. These assays can be used to measure reactions that are impossible to assay in any other way.[6]Chemiluminescent[edit]Chemiluminescence is the emission of light by a chemical reaction. Some enzyme reactions produce light and this can be measured to detect product formation. These types of assay can be extremely sensitive, since the light produced can be captured by photographic film over days or weeks, but can be hard to quantify, because not all the light released by a reaction will be detected. The detection of horseradish peroxidase by enzymatic chemiluminescence (ECL) is a common method of detecting antibodies in western blotting. Another example is the enzyme luciferase, this is found in fireflies and naturally produces light from its substrate luciferin. Light Scattering[edit]Static light scattering measures the product of weight- averaged molar mass and concentration of macromolecules in solution. Given a fixed total concentration of one or more species over the measurement time, the scattering signal is a direct measure of the weight- averaged molar mass of the solution, which will vary as complexes form or dissociate. |