• Identification and characterization of CRIP1b: a novel CB₁ cannabinoid receptor interaction protein

      Niehaus, Jason Lance; School of Graduate Studies (2006-07)
      G protein-coupled receptors. (GPCRs) transduce extracellular stimuli to intracellular signals. through their interaction with heterotrimeric G proteins. Signaling diversity and specificity is imparted primarily through variations of G protein subunits. Protein-protein interactions between intracellular accessory proteins and GPCRs also modify signaling by altering receptor activity or signaling pathways. The ability of intracellular proteins to interact with the CB 1 cannabinoid receptor was investigated to determine whether particular signaling properties of CB 1 resulted from interaction with specific CB 1 interacting proteins. A novel protein named CRIP 1 b was discovered· to interact with the C-terminal tail of CB 1 ~- The interaction between CRIP 1 b and CB 1 was characterized using the yeast two-hybrid assay. Functional consequences of the CRIPlbCB 1 interaction were investigated by examining protein localization by confocal microscopy and measuring CB1 mediated N-type Ca2+ channel activity in the presence of CRIP 1 b by whole-cell patch clamp recordings. The ye_ast two-hybrid assay indicated that the last nine amino acids of the. CB 1 C-terminal tail were required for interaction with CRIP 1 b. Heterologous expression of CRIP 1 b and CB 1 in HEK 293 cells did not reveal evidence of co localization, nor was CB 1 able to significantly traffic CRIP 1 b to the plasma membrane. However, CRIP 1 b and CB1 were found to colocalize in superior cervical ganglion (SCG) neurons. Whole-cell voltage-clamp recordings ofN-type Ca2+ channels in SCG neurons indicated that CRIPlb had no.effect on agonist- or inverse agonist-induced modulation of Ca2 + current by CB1. Furthefrr1:ore, the level of CB 1 constitutive activity was· not significantly altered by CRIP 1 b. The high affinity of CB 1 for G proteins, as demonstrated by the ability of CB 1 to sequester G proteins from other Gu0 coupled receptors, was unaffected by expression ofCRIPlb. These results provide evidence that CRIP 1 b is a novel CB 1 accessory protein that interacts with the C-terminal tail of CB1. While CRIPlb and CB1 can apparently interact in a neuronal expression system, the ability of CRIP 1 b to modify CB 1 signaling was not detected in any of the pathways investigated. Thus, the distinctive signaling properties of CB1, such as constitutive activity and G protein seque.stration do not originate from nor are modified by CRIPlb.
    • Identification and Characterization of Force Sensitive Domains using an In Vivo Dorosphila Synthetic Biology Platform

      Harman, Jacob Henry; Biomedical Sciences
      The Notch protein is a family of highly conserved transmembrane signaling proteins that are found in all metazoan life. This protein family plays many roles in developmental and regulatory pathways in these organisms such as the development of the human vertebral column and the Drosophila adult wings. The Drosophila Notch protein has been the focus of recent research as this protein has shown to be dependent on a unique signaling mechanism that relies on the force being applied to the receptor by the endocytosis of a bound ligand for a cleavage event to occur. This protein has been shown to only signal in the presence of this endocytosis force which allows for cleavage at the S2 and S3 sites on the protein which allows for a transcription factor to be released from the membrane and enter the nucleus. In synthetic biology, this protein has found a role in creating customizable cell-cell signaling platforms known as Synthetic Notch or Syn-Notch. As demonstrated in previous research, Notch proteins contain highly modifiable ligand binding and transcription factor domains that can be interchanged with other ligand binding and transcription factor domains from non-Notch proteins without affecting the protein signaling. In addition to the ligand-binding and transcription factor, the Notch protein contains a Negative Regulatory Region (NRR) which functions as a force sensor domain and allows for the force signaling activation seen in the native Notch proteins. This region of the Notch protein is often conserved in Syn-Notch systems as a means of controlling the signaling of Syn-Notch proteins used in precision medicine. The unique force-sensing function of this domain has raised questions regarding the characteristics of this domain that allow for it to act in a force-sensing manner and if this domain is as interchangeable as its other components. In this study, domains from other non-Drosophila Notch proteins and non-Notch protein domains were evaluated in an in vivo Drosophila platform for the ability to function as a force sensor domain. In this screen, several unique force sensor domains were identified as having the ability to recapitulate the force sensing characteristic of the canonical Notch NRR. While a common thread between these domains in terms of defining characteristics remains elusive, this study was able to demonstrate the feasibility of this screening method to identify force-sensitive domains and identify 4 domains that exhibit this characteristic. The identification of these domains which are force sensitive not only allows for the characteristics to function as force-sensitive domains but also provides alternative force sensors with differing force sensitivities when compared to the canonical Notch NRR for use in Syn-Notch systems. The development of these receptors will not only contribute to the field of synthetic biology as means to create synthetic platforms for use in future research, but these receptors will allow for the development of even more precise treatments using these synthetic receptor systems in clinical therapies.
    • Identification of insulin-responsive genes in rodent bone cells

      Blythe, Gregory A.; Department of Oral Biology (Augusta University, 1998-06)
    • Identification of Muscarinic Receptor Subtypes and Arachidonic Acid Metabolites Mediating the Effects of Actylcholine in The Pulmonary Circulation of Rabbits and Cats

      El-Kashef, Hassan; Department of Pharmacology (1987-05)
      Acetylch.oline (ACh) is known ·to reduce vascular pressure throughout the systemic. circulation; in. the·-- pulmonary circulatfon: ACh: can·- produce:either vasodi-1ationr or' ·vasoconstri-ction. For example,. ACh', produces: vasoconstrictfon · in· the, pulmonary ·circula~i on· of rabbits .. but dtlati on in· the· -feline. pu·lmof!ary-, vasculature·~-- The·· mechanism(s-) responsible·' for· the· actions of ACh in the pulmona.ry vasculature-- remain unc;lear. This study was designed to i·nve·st.igate poss·ible mechanisms. responsible for the actions. of ACh in the ra~bit ancf cat pulmonari_.circulation·~- Our hypothesi's is tbat . variations in· the· v.as.cular ·-response~ to: ACh may- be due to differe.nces in med-iators. (e.g. ·prostanoids)· released upon. activation .. of the muscarinic receptor~ These· di.ffe-rences cou 1 d be attributed,· either to. differences ... in muscarinic receptor subtypes.-. or to differences. in the·. metabolism of· arachidonic acid- tn rabbit. vs. cat. In anesthetized rabbits,_ ACh-induced pulmonary=- vasoconstriction-··. was,. totatly, inhibited::'·- by:: the: phosphotipasey A'2· inhibitor· qu-inacrine··,. the cycl o~oxygenase"' inhibitors:; indomethacin·,. and,.~ mec lofenamate·,, the- thtomboxane/ A:2:· synthetase=, i nh~i bi:tot· ]-;.;.{1'- Imidazolyl.)-· Heptano.fc. Acid .. (7;..IHA), and'- by,- the··' thromboxane·· A- 2 . receptor·· antagpntst. SQ:. 29-~.548:, .. but not·,. by·- the· 1 .ipoxygenase:-- inhibitor- nordihydrogua-iaret.ic: acid( NDGA}':. AGh·: s:igni-ftcantly; increased> plasma; thromboxane···B2.' (TXB'2J:· levels;. in·-. anesthetized:·,_ rabb_its:. A.Ch:~induced:. decrease'~ in, system.ic ar.teria:l pressure'" was, not· affected: by:·. any o.f'· the·· i nh'ibftors,. mentionedJ above--._ Small>. doses: of· the, select.i ve:· Mi- muscari nfc. receptor· subtype· antagonists:, pi renzepihe and_:. tr.ihexypheni:d~l,.. s:i gni-f.i cantly:- fnhi bjted:-: the· ACh.-i nduced:~- increase:'-·. i m pulmonary; vascuJa·r res:fstance- but. not: the~ ACh-induced=;· decrease Ht systemic. arterfal~: pressur.e··. The:.- s·ame" dose·· o.f secover.i ne:, .. a-~.- se·l ecti ve, M2-- muscar.i.nic: ii recept.or subtype. antagonist, dld ·not· change the · ACh ·effects on the pu 1 mona ry or· sys.temi c · v qSCu latu,re. Larger doses of pirenzepine, trihexyphen-idyl and\· secoverine,· totally. inhibited· the.- effects:: of ACh in· the .. pu:lmonary· and: systemic: circulatjons. Atrop.ine··. equally:- inh.i.bi ted:· the· effects. of ACn· on'" the pulmonary..- and.: systemic vascul atures- at.- any. do$es~ . . used': In .. fso_lated· rabbit·. lungs .. perfused·:·in:· situ~: w:ith · b]ood·-,_'free· medium;< . . . . . . . the ACh-fnduced· increases fn .. ·pulmonary v.ascular resistance, TXB2 level·s: a:nd ·6-Keto~PG·Fra were significa.nt.ly inhibited. by small concentrations.· of· pi.renzepine but .not· secove·rine. Larger concentrations. of· p.irenzepine~ and:'· · . . secover·ine totally ·inhibited· these· effects of ACh. In -anesthetized c~ts, the· ACh~inducec:l decre-ase in pulmonary vascular resfstance was partially~ · inhibited· by quinacrine and·' indomethacin and· significantly:· .potenti~ted by NOGA. The ACh-induced ·decrease in systemic arterial .pressure was not s·ignificantly affected by· any. of these· inhibitors. Small. doses. of secover-ine:'' but not . pfrenzepi ne;~" i nh.ibited·· the: pulmonary: but,. not .. the:· systemic vascular-. response·, tct.. ACh.,,_ i!!.=- v:ivo~ Howev-er,_. large··· doses; .. of·.· ptrenz:epi ne-, · tri·hexyphen:fdyl. and' s.ecoveri'ne' significantly:· inh.i bited~ the:, .. . . effects. of:. ACh~ in·.: the~: pulmonary;· and. systemic vascu:l ature\ In: isola ted:· cat: 1 ung_s:. perfused~- fm: sitw: w.ith; bJ ood.;.free- · medium~. 'ACh'" di-lated... the·· precons.tricted:: pulmonary: artery. bu.t> tt: did·: not ·s,tgnific.antly:· alter- TXB2 ori 6~ Keto~PGF 1 at 1 eveTs:~.. Smal:T: concentrat:i ons:- of:-· secoverfne-': but· not· pJ.renzep.i ne~-- par.t.ially;' i nhi bi'tedr the· ACh;.. induced~· decrease· in: pulinonar,Yvascular ·· resistance· •. The· pros.tacyc;ti n .. · synthetase fnhi bitor· tranylcypromine· si"gn.f$icantly~ i'nhi bited;'; the,· ACh- induced§ decrease:· in:· · pu]monary.:; va-scu..lar· resi.stance·~- . . i fi: ·These· results. ·indi-cate that l) ACh has . opposite actions in. the systemic {dilatory) versus pulmonary· (constrictor) circulation of rabbi-ts, 2r Arachidon-ic· acid.'metabo'lftes·. mediate. the-. pulmonary: . but. :not·· .the" systemic vascular respon.se- to. ACh, in rabbit, 3}. Thromboxane·· A2 mediates,. the:· pulmonary- vasoconstrictor- response·· to, ACh in rabbit·~ 4} The r.abbit pulmonary . v~scular· muscarinic recepto.rs. are· very·,· sensit:ive to·. p_irenzep.i-ne· . and thus be:have m9re · 1 ike Mi recept~rs, _5) · In· the cat·, the· ACh- induced·· decrease in- p~lmonary but not systemic vascular. pressure is partly mediated by prostacycli'n, 6)' In _the cat, the vascula-r· muscarinic receptors both in the· pulmon~ry. and the systemic beds are not selectively sensitive to . .pirenz.ep.ine ·and- thus behave like. non-Mt receptors and,· 7) In the .cat, the· ·muscarinic receptors. in the· pulmonary. and, systemic vasculat~re· represent different or heterogenous populations of M2. receptors since· they exhibit different affinities·. to secoverine.
    • Identification of novel molecular biomarkers for diagnosis of salivary dysfunction

      Suart, Mary S.; Medical College of Georgia (Augusta University, 2013-10)
    • Identification of professional competencies needed for practicing registered nurses in urban and rural hospitals

      Isler, Barbara A.; School of Nursing (1981-05)
      The purpose of this study was to determine if the hospital- location .s i gni fi cantly influenced the regis ter.ed nurses• competency needs .. A. sample of 112 urban and 80 rural .nurses was selected by Nursing Directors in 23 Georgia hospitals. Data were collected by use of a 113 item . competency questionnaire adapted from Clayton'·s (1978) work in thi.s area .. · Findings suggest the urban o~ rural hospital setting significantly . influences the competencies used by the registered nurse. Other findings include the acceptance percentage for ea-ch competency by the urban or. rural nurse indicating thei~ perception of nee~ and demographic characteristics of the two nurse populations.
    • Identification of Regulatory Elements in a Conserved Upstream Region of the Gene Encoding Interphotoreceptor Retinoid-Binding Protein (IRBP)

      Lu, Haiyan; Department of Ophthalmology (1999-06)
      (First Paragraph) IRBP is a large, single-subunit extracellular glycolipoprotein found in the interphotoreceptor matrix between the photoreceptor cells and retinal pigment epithelium cell layer (Fig. 1.). The protein is synthesized and secreted by the photoreceptor rods and cones, as well as pinealocytes, of all vertebrates. The molecular weight of human IRBP (1230 residues) is 133,400 daltons. This protein consists of four homologous segments of approximately 300 residues each. Each segment contains highly conserved hydrophobic domains among species. Ligands identified as bound to IRBP include retinoid isomers and fatty acids, and IRBP can also bind cholesterol, a-tocopherol and retinoic acid. The ability of IRBP to bind various retinoid isomers, fatty acids and many other hydrophobic ligands suggests multiple functions in the retina .
    • The Identification of the Requisite Knowledge and Relevant Role Activities by Nurse Administrators and the Relationship of These to Their Educational Preparation and Management Level

      Allen, Lori; Department of Nursing (1988-01)
      The purpose.of this descriptive correlational study was to identify the requisite knowledge and relevant role activities needed by nurse administrators to successfully enact their roles (first, mid, and top) and the relationship of these role knowledge areas and role.activities to their educational preparation and management level. To achieve the study purpose, two research questions and three hypotheses were tested. A total of 226 subjects participated in the study: 156 first-line nurse administrators; 44 mid level administrators.; and 26 top level nurse administrators. The study population consisted of a convenience sample of nurse administrators from five hospitals in a Southeastern metropolitan area. Data were collected through the use of the Nurse Administrative Role Aetivities ·and Knowledge (NARAA~) tool. Descriptive statistics were utilized to address the first research question. To test the hypptheses related to the second research question, ANOVA were utilized-. All three hypotheses were supported at the p=.05 level. More specifically, 13 role activities and four requisiteknowlege areas were found to be significantly different for first level .. and .top level administrators. The majority of these items were related to leadership, management, and administration of human resources. Seven role activities and 11 requisite knowledge areas were found to differ on the basis of educational preparation. Nurse administrators at each organizational level identified significantly different role activities·relevant for.their successful role enactment, but did not identify different requisite role knowledge areas. There was no significant difference in how nurses with different academic degrees rated the relevance of role activities to the success of their role enactment on the basis of academic degree. Academic degree did, however, discriminate among respondents in the degree to which they rated the relevance of requisite knowledge areas to the success of their role enactment. Other findings, limitations, implications, and suggestions for further study were discussed.
    • Identifying a vascoconstrictor role for interleukin-6, a pro-inflammatory cytokine.

      Daniels, Ericka R.; Medical College of Georgia (Augusta University, 2004-05)
      Increased plasma levels of interleukin-6 (IL-6) have been associated with elevated blood pressure and is predicative of risk for a firSt myocardial infarction. We therefore hypothesized that interleukin-6 has vasoconstrictive properties. Methods for this study included quantification of tissue levels of IL-6 , by capture ELISA from angiotensin 11- . I i hypertensivie male, C57BL/6J mice. Ti~sue coJcentratioti of IL-6 was compared to con~rol normotensiv¢ samples (thoracic aortic segments). ·Blood pressure was monitored by telemetry and the direct effect of IL-6 on v.ascu~ar contractility was studied in thoracic aortas isolated from normotensive mice. Our results show that IL-6 was significantly elevated (231.4 pg/mg ± 13.4 pg/mg . . ~ . . . SEM) in angiotensin II-hypertensiye mice compared to controls (129.9 pg/mg ± 11.2 · pg/mg SEM) and WT mice displayed a higher increase in MAP to angiotensin 11-infusion (60.35 mmHg ± 4.55 mmHg) at the end of infusion period compared to IL-6 KO mice (33.81 mmHg ± 6.67 mmHg). Pre-treatment of denuded aortic rings from normotensive mice with IL-6 for 24 hours caused a significant inhibition of their contractile response to phenylephrine and endothelin-1. In contrast, there was no significant effect on relaxation response to sodium niroprusside. In endothelium intact aortic rings, IL-6 caused contraction in the presence of L-NAME to inhibit nitric oxide synthase. We conclude that depending upon the experimental conditions, IL-6 has vasoconstrictive properties.
    • Identifying Challenges and Solutions to Mentoring in a Rural School District

      Boyington, Justin Keith; Department of Advanced Studies and Innovation (Augusta University, 2021-05)
      The purpose of this study was to investigate the experiences of mentors and mentees in the induction program in the rural county of Greenway county school district. Mentors and mentees were able to share their experiences in the induction program through interviews, questionnaires, and an audio reflective journal. Mentors specifically identified their role as a mentor within the induction program and how it contributed to new teacher efficacy in the classroom. Additionally, mentors and mentees were able to share challenges in the induction program such as excessive workload and mismatching between mentors and mentees. Finally, mentors had the opportunity to identify possible solutions to help address these challenges and create a more effective induction program for years to come. The findings of this study provided insight on strategies to improve induction programs including providing virtual supplements for mentors and mentees and involving mentors in decision making within the induction program.
    • Identifying Challenges and Solutions to Mentoring Teachers in a Rural School District

      Marshall, Carlos Jimmie; Department of Advanced Studies and Innovation (Augusta University, 2021-05)
      The purpose of this study was to identify perceived challenges that mentor and mentee teachers face within the teacher induction program in the rural school district of Greenway County. The different perceptions of mentor and mentee teachers will provide rich insight regarding the challenges teachers face within these programs. The challenges presented by the mentor and mentee teachers will help to identify areas within the induction program that require additional assistance or possible reform. This study is intended to propose possible solutions suggested by the mentors to address the challenges identified. This qualitative research study utilized various methods of data collection to investigate the lived experience of mentor and mentee teachers in the induction program. Data was collected from mentors using an online questionnaire, a semi-structured interview, and reflective journals, while a focus group interview was utilized to collect data from mentees. The survey questionnaire helped to provide context to the lived experience as a mentor teacher in the induction program within Greenway County. Semi-structured interviews with the mentor teachers allowed for the emergence of possible solutions to best address identified challenges resulting in the overall effectiveness of the induction program. Audio reflection journals were collected after mentor-mentee meetings to gather a deeper understanding of mentor-mentee interaction, while also deepening the understanding of challenges that arose during the induction program. Finally, data was collected from mentee teachers through a focus group interview to gain their first-hand experience of the induction program and their mentor’s role within the program. Keywords: Mentoring, mentors, mentees, induction programs, challenges
    • Identifying Challenges and Solutions to Mentoring Teachers in a Rural School District

      RALSTON, KRISTINA MIZE; Department of Advanced Studies and Innovation (Augusta University, 2021-05)
      Approximately 50% of new teachers leave the profession of education within their first five years (Burton & Johnson, 2010; McCoy, 2019; Ingersoll & Smith, 2004; Steinke & Putnam, 2011). With such a mass exodus of teachers, schools should develop programs to promote new teacher retention (Callahan, 2016; Kent et al., 2012). Induction programs offer new teachers support by providing them with a mentor teacher to support new teachers' needs (Ingersoll & Smith, 2004). However, to offer such beneficial programs to new teachers, schools must attract experienced teachers to fulfill a mentor's role. The research team investigated a rural county’s induction program to evaluate the challenges mentor teachers face. Mentor teachers offered their suggestions as possible solutions for the improvement of the induction program. These proposed solutions could be used to improve the quality of induction programs which may promote teacher retention.
    • Identifying the Function of Nasal Embryonic LHRH factor (NELF) in Immortalized GnRH Neurons

      Ko, Eun Kyung; Department of Neuroscience and Regenerative Medicine (7/30/2018)
      Hypothalamic gonadotropin releasing hormone (GnRH) is crucial for the proper function of the hypothalamic-pituitary-gonadal (HPG) axis, puberty, and reproduction. When GnRH neuron migration or GnRH regulation is impaired, hypogonadotropic hypogonadism results. Mutations in the gene for nasal embryonic LHRH factor (NELF) have been identified in GnRH-deficient humans. NELF is a predominantly nuclear protein that may participate in gene transcription, but it is unlikely to be a transcription factor. Thus, our hypothesis is that NELF may indirectly regulate transcription via protein-protein interaction within a transcription complex. To address this question, RNA was extracted from NLT GnRH neuronal cells following either stable Nelf knockdown or scrambled control and subjected to cDNA arrays. Expression of transcription factors and cell migration gene expression was most commonly altered. Members of the Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway, including Stat1, Stat2, Stat5a, Jak2, Irf7 and Irf9, were significantly down-regulated as assessed by RT-qPCR. Protein levels of STAT1, phospho-STAT1, and JAK2 were reduced, but the levels of phospho-JAK2 were not. These findings suggest a role for NELF in the regulation of the JAK/STAT signaling pathway, which has important functions in GnRH neurons. Jacob, the rat orthologue of NELF, is phosphorylated at the serine 180 residue by phosphorylated Erk1/2 which is activated by synaptic NMDA receptor activation and then translocates to the nucleus. Phosphorylated Jacob in the nucleus interacts with CREB and regulates the expression of BDNF, which is associated with synaptic plasticity in the brain. Proteins, such as caldendrin, importin-α and α-interxin, have been identified as binding proteins with Jacob. However, binding proteins, phosphorylation sites and/or kinases for NELF have not yet been reported. To demonstrate novel and putative functions of NELF in the nucleus, identification of binding proteins and phosphorylation sites is required. To address this question, co-immunoprecipitation (Co-IP), mass spectrometry and in vitro kinase assays were performed. We found six putative binding proteins that could interact with NELF, including 14-3-3ε. We also identified phosphorylation sites on NELF, including serine 288, which could be phosphorylated by AKT1 kinase. These new findings will be helpful to understand the function of NELF in the nucleus
    • Immediate healing responses to ceramic endosseous dental implants in dog gingiva

      Long, William Gregory; Department of Oral Biology (1990-05)