The p16INK4a tumor suppressor is upregulated and serves to limit proliferation in response to cellular stressors and oncogenic events. Induction of p16INK4a is highly dynamic during tumorigenesis and aging, and is expressed in response to a variety of oncogenic stressors. The novel knock-in mouse model, p16-LUC, wherein the endogenous p16INK4a promoter drives single-copy expression of firefly luciferase, is an excellent marker of tumorigenesis and aging. p16 induction can be monitored dynamically in vivo following treatment with DNA damaging stressors such as ionizing radiation or can serve as a biomarker in studies investigating oncogenic stress, oncogenic drivers, early tumorigenesis, novel anti-neoplastic therapeutics, toxicities, senescence, and other in vivo stress responses.  This model is available on an albino C57BL/6 background (haired). The conditional knock-out mouse model, p16-floxed, contains loxP sites in the Cdkn2a gene before the exon 1a start site (left) and in the first intron (right), such that only p16INK4a and not p19ARF is affected by removal of exon 1a. This strain can be crossed with Cre-expressing strains to investigate how the loss of p16INK4a affects early oncogenesis and senescence.  This model is on a mixed C57BL/6 and 129 background. Please contact Dr. Shenghui He at shenghui@email.unc.edu or Steve Pedroza at spedroza@email.unc.edu to inquire about availibility of this research tool.

The P2Y1 protein has been shown to help in the formation and stabilization of the thrombus. P2Y1 deficient mice exhibit longer bleeding times and are useful in the validation of prothrombotic and antithrombotic therapies. These mice are available from The Jackson Laboratory and can be found at https://www.jax.org/strain/009131. 

EP3 is a receptor for prostaglandin 2. The EP3 receptor has been linked to prostaglandin-related processes such as pain, tumor growth and inflammation. These mice can be used to investigate the effects of EP3 modulators. If you are an academic institution or nonprofit organization interested in this research tool for noncommercial purposes, please contact the researcher directly to inquire about availability.

TDP-43 proteinopathies including frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS) are neurodegenerative disorders characterized by aggregation and mislocalization of the nucleic acid-binding protein TDP-43 and subsequent neuronal dysfunction. This mouse model is an endogenous model of sporadic TDP-43 proteinopathy based on the principle that disease-associated TDP-43 acetylation at lysine 145 (K145) alters TDP-43 conformation, impairs RNA-binding capacity, and induces downstream mis-regulation of target genes. Expression of acetylation-mimic TDP-43K145Q results in stress-induced nuclear TDP-43 foci and loss of TDP-43 function. Mice harboring the TDP-43K145Q mutation recapitulated key hallmarks of FTLD, including progressive TDP-43 phosphorylation and insolubility, TDP-43 mis-localization, transcriptomic and splicing alterations, and cognitive dysfunction. This mouse is available from the Mutant Mouse Resource and Research Centers (MMRRC) Cat #: MMRRC 068119-UNC TDP43KQ (C57BL/6J-Tardbpem1Toco/Mmnc). https://www.mmrrc.org/catalog/sds.php?mmrrc_id=68119

BALB/c mice are commonly used for B cell hybridoma development because of their predictable B cell responses to various antigens. Our investigators have developed the tg801 transgenic mouse line, (BALB/c latency mice), as an alternative to the commonly used BALB/c or BALB/c/FcgR2b KO mouse that are often used for studying B cell immunobiology. These BALB/c latency mice express genes from the Kaposi sarcoma-associated herpesvirus that are associated with the long-term latency period of infection. These mice develop hyperglobulinemia, plasmacytosis and B cell hyperplasia. When these mice are exposed to viruses, such as Zika virus, they exhibit enhanced antibody response and infection clearance relative to wild type BALB/c mice. These mice are ideal for studying B cell immunobiology and production of monoclonal antibodies. If you are an academic institution or nonprofit organization interested in this research tool for non-commercial purposes, this mouse is available from Jackson Laboratories (Stock No. 032639) https://www.jax.org/strain/032639  

This virtual educational seminar series was created by clinicians from the UNC Adams School of Dentistry. The seminars are well suited to complement the coursework for anesthesia with nitrous oxide or benzodiazepines that are taught in Dental Hygiene, Dental Assisting, Dental Student, Medical Student, or Clinical Graduate Training programs. The individual seminars can be viewed alone or in combination. The seminar topics and the learning objectives for each are as follows: Seminar One: Depths of Sedation/ Anesthesia and the Central Nervous System How anesthesia evolved  Differentiate among the changes in the CNS with sedation/anesthesia drugs Seminar Two: Routes for Drug Administration, Pharmacology of Nitrous Oxide, and Benzodiazepines Compare sedation and general anesthesia by physiologic outcomes  Compare drug administration by access, control of dosage  Relate drug administration to elimination; alpha half-life (redistribution) and beta half-life (elimination)  Describe possible clinical outcomes associated with choice of drug administration Seminar Three: Pulmonary/Respiratory Physiology: A Review Discuss relationship between lung function and gas exchange at the alveolus  Discuss relationship between monitoring with pulse oximetry and arterial oxygen levels  Compare monitoring: pulse oximetry vs. capnography   Describe how sedative drugs might affect blood oxygen levels Seminar Four: Evaluation of patients for sedation/anesthesia Classify patients by health status Assess patients specifically as candidates for inhalation anesthetics Seminar Five: Anesthesia / Monitoring Equipment Sequence technical steps for administration of N2O  Select monitoring protocol for individual patient  Compare clinical uses of pulse oximetry and capnography Seminar Six: Administration of Nitrous Oxide Prepare patient for planned anesthesia: diet, caregiver responsibility  Decide levels of anesthesia based on the patient’s responses  Sequence steps if patient becomes agitated, disoriented, nauseated  Sequence steps if airway blocked  Implement discharge protocol after anesthesia Seminar Seven: Side Effects / Hazards to Staff of Nitrous Oxide Evaluate patients for high risk of nausea/vomiting  Evaluate patients for high risk of vitamin B12 deficiency  Explain hazards to staff  Understand mechanisms of reducing N2O in ambient air A preview of the seminar series can be found by downloading the preview document on the right. This preview contains links for viewing the Introduction and Seminar One videos, as well as a course outline, and learning objectives, references, and an evaluation for Seminar One. The seminar series is available for licensing for educational use. All the material needed for implementing the seminar series in a course is provided (seminar videos, course outline, learning objectives, and seminar evaluations). The seminar material, which contains links for streaming the seminar videos, will be available for download following acceptance of the terms of the license agreement and payment of the license fee.   Offerings Add Offering Name                                                                                                                 Price           Shipping    Currency Nitrous Oxide_Benzodiazepines Anesthesia - 21-0069                                    $500.00         $0.00           USD Demo Nitrous Oxide/Benzodiazepines Anesthesia - 21-0069                           $0.00             $0.00          USD

10-0081 FLUOROGENIC REPORTER DETECTS PHOSPHOLIPASE C ACTIVITY IN MAMMALIAN CELLS Benefits: Provides quantitative data of PLC isozyme activity in mammalian cells without radiolabeling Facilitates high throughput screens for PLC inhibitors, enabling identification of novel PLC-inhibitory compounds Identify time and location of PLC activity by linking reporter to cellular localization signal, photocage, and/or quencher Phospholipase C isozymes (PLC) are major signaling proteins that function in a vast range of cellular processes. PLC dysregulation is implicated in diverse health conditions including cancer, Alzheimer’s, epilepsy, arthritis, and neuropathic pain, making PLCs attractive targets for pharmaceutical interventions. Understanding the intricacies of PLC regulation requires specific reporters that identify a cell’s PLC activity and precise inhibitors that dissect the role of PLC in a biological process. However, methods to measure PLC activity require radiolabeling or work only in bacteria, and the few extant PLC inhibitors cause pleiotropic effects. A novel fluorogenic reporter developed by researchers at The University of North Carolina at Chapel Hill, small molecule WH-15, offers considerable improvements: it directly monitors PLC activity in mammalian cell lysates, enabling quantification without radiolabeling, and it can be used in high throughput screens to identify new inhibitors. WH-15 has an inositol phosphate headgroup like PLC’s endogenous substrate, membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PIP2), but features a linker that bridges the headgroup to the fluorophore 6-aminoquinoline. PLC activity cleaves WH-15, forming IP3, a quinomethide byproduct, and the fluorescent molecule 6-aminoquinoline. WH-15 is not fluorescent, but cleavage by PLC activity releases 6-aminoquinoline, increasing reaction fluorescence over 15-fold to produce a signal that is easy to measure with 355 nm excitation/535 nm emission. PLC affinities for WH-15 and its endogenous target are similar, indicating that WH-15-released fluorescence accurately reflects PLC efficiency. Importantly, WH-15 is general enough to work with diverse PLC isoforms but is specific to active PLCs—PLD, PLA2, and engineered catalytically dead PLC did not increase reaction fluorescence. The research team used WH-15 to rapidly identify and validate three new PLC inhibitors in a screen of a 6280-compound library. WH-15 can be modified to replace the headgroup with any PLC substrate, change the linker, and/or add cellular localization signals, photocages, and/or quenchers to study specificity. WH-15 is stable at room temperature for up to 24 hours. These features make novel reporter WH-15 useful for measuring PLC activity in different cell types and disease states and for identifying potential PLC inhibitors in high throughput screens. Related publications: Huang, W, SN Hicks, J Sondek, and Q Zhang. 2011. A Fluorogenic, Small Molecule Reporter for Mammalian Phospholipase C Isozymes. ACS Chem Biol 6:223-228. https://pubmed.ncbi.nlm.nih.gov/21158426/ Huang, W, M. Barrett, N Hajicek, S Hicks, TK Harden, J Sondek, and Q Zhang. 2013. Small Molecule Inhibitors of Phospholipase C from a Novel High-throughput Screen. J Biol Chem 288(8):5840-5848. https://pubmed.ncbi.nlm.nih.gov/23297405/

Although hepatitis C virus (HCV) is a leading cause of cirrhosis and liver cancer, many details of its replication remain obscure, as most HCV strains replicate poorly in cell culture. Furthermore, no effective vaccines are available to date and the mechanism of HCV persistence and carcinogenesis in vivo remain inadequately understood. This plasmid contains infectious molecular clones of H77D virus, a replication competent genotype 1a variant of HCV. These clones are capable of robust replication in cultured cells and could be used to facilitate drug development and the assessment of viral resistance to therapeutics. This plasmid is available for distribution from Kerafast and can be found at https://www.kerafast.com/product/3453/.

Abstract G protein-coupled receptors (GPCRs) are essential mediators of cellular signaling and are important targets of drug action, with over one third of prescribed medications targeting the non-olfactory class of these receptors.  GPCRs have a large diversity in their signal transduction cascades necessitating the use of numerous functional assays to identify agonists or inhibitors of these receptors. Assessment of GPCR-ß-arrestin interactions offers a way to universally assay a large variety of GPCRs. Transcriptional activation following arrestin translocation (TANGO) assays allow for effective high throughput screening of GPCR activity by assessing ß-arrestin-GPCR activity independent of coupling of ß-arrestin to a G-protein. This allows for interrogation of so-called “orphan” GPCRs for whom the receptor’s ligand and/or its coupled G-protein may not be known.   Researchers at UNC have developed open-source resource that provides the capacity to interrogate the druggable human GPCR-ome via a G-protein-independent ß-arrestin recruitment assay. This method is termed is termed PRESTO-TANGO (Parallel Receptor-ome Expression and Screening via Transcriptional Output - TANGO). Researchers have applied this unique platform to more than 120 nonorphan human GPCR targets, and demonstrate its utility for discovering new ligands for orphan human GPCRs.     These PRESTO-TANGO plasmids are available for academic institutions or nonprofit organizations interested in utilizing this research tool for noncommercial purposes, at Addgene (Roth Lab PRESTO-Tango GPCR Kit, Kit # 1000000068). https://www.addgene.org/kits/roth-gpcr-presto-tango/   Benefit ·       The PRESTO-Tango Kit can be used to measure receptor activation for more than 300 GPCRs via a modified Tango beta-arrestin recruitment assay. ·       Facilitates the rapid, efficacious, parallel, and simultaneous profiling of biologically active compounds across essentially the entire human druggable GPCR-ome. ·       PRESTO-TANGO is sensitive, easily executed, and amenable to both HTS and simultaneous parallel screening at many GPCRs. ·       Particularly suited for “first-pass” screening of compound libraries for identifying ligands of orphan receptors, since knowledge of the G protein partners of each GPCR is not required  

Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that causes outbreaks of debilitating acute and chronic arthralgia in humans. There are currently no approved vaccines and few therapies against CHIKV. This antigen preparation is derived from a full length molecular clone based on a contemporary clinical CHIKV isolate and can be used to generate diagnostic reagents against CHIKV. Inactivated whole virion CHIKV antigen has the potential to screen patient or animal serum for the presence of CHIKV specific antibodies, act as an antigen for generating CHIKV specific monoclonal antibodies, or produce an inactivated vaccine against CHIKV. If you are an academic institution or nonprofit organization interested in this research tool for noncommercial purposes, please contact the researcher directly to inquire about availability.