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Biomedical subjects

Richard N Re

Publications and source records attributed to Richard N Re.

18 recordsLinked to original sources
PubMed2006

Cleavage of the angiotensin II type 1 receptor and nuclear accumulation of the cytoplasmic carboxy-terminal fragment.

Our published studies show that the distribution of the ANG II type 1 (AT(1)) receptor (AT(1)R), expressed as a enhanced yellow fluorescent fusion (YFP) protein (AT(1)R/EYFP), is altered upon cellular treatment with ANG II or coexpression with intracellular ANG II. AT(1)R accumulates in nuclei of cells only in the presence of ANG II. Several transmembrane receptors are known to accumulate in nuclei, some as holoreceptors and others as cleaved receptor products. The present study was designed to determine whether the AT(1)R is cleaved before nuclear transport. A plasmid encoding a rat AT(1)R labeled at the amino terminus with enhanced cyan fluorescent protein (CFP) and at the carboxy terminus with EYFP was employed. Image analyses of this protein in COS-7 cells, CCF-STTG1 glial cells, and A10 vascular smooth muscle cells show the two fluorescent moieties to be largely spatially colocalized in untreated cells. ANG II treatment, however, leads to a separation of the fluorescent moieties with yellow fluorescence accumulating in more than 30% of cellular nuclei. Immunoblot analyses of extracts and conditioned media from transfected cells indicate that the CFP domain fused to the extracellular amino-terminal AT(1)R domain is cleaved from the membrane and that the YFP domain, together with the intracellular cytoplasmic carboxy terminus of the AT(1)R, is also cleaved from the membrane-bound receptor. The carboxy terminus of the AT(1)R is essential for cleavage; cleavage does not occur in protein deleted with respect to this region. Overexpressed native AT(1)R (nonfusion) is also cleaved; the intracellular 6-kDa cytoplasmic domain product accumulates to a significantly higher level with ANG II treatment.

Angiotensin II↗
PubMed2006

Nuclear accumulation of the AT1 receptor in a rat vascular smooth muscle cell line: effects upon signal transduction and cellular proliferation.

The objective of the study was to identify the functional outcome of intracellular versus extracellular angiotensin II-AT(1) receptor interactions in vascular cells. Rat vascular smooth muscle cell line A10 was transfected, independently and concurrently, with plasmids encoding fluorescent fusion proteins of rat angiotensin II (pECFP/AII, encodes AII fused downstream of enhanced cyan fluorescent protein) and the rat AT(1a) receptor (pAT(1)R/EYFP, encodes the rat AT(1a) receptor fused upstream of enhanced yellow fluorescent protein). The AII fluorescent fusion protein possesses no secretory signal peptide and deconvolution microscopy established that is maintained within these cells predominantly in the nucleus. AT(1)R/EYFP was absent from the nucleus when expressed exclusively or in untreated cells but accumulated in the nucleus following exogenous AII treatment or when co-expressed with ECFP/AII. Furthermore, expression of ECFP/AII stimulated proliferation of A10 vascular smooth muscle cells (VSMCs) 1.6-fold (P < 0.05). Transfection of a control, pECFP/AII(C) (which encodes a scrambled AII peptide fused to ECFP) had no growth effect. In light of the intracellular growth effects of ECFP/AII, we sought to elucidate the underlying signaling pathways. We found that extracellular AII treatment of A10 cells activated cAMP response element-binding protein (CREB) as determined by one-hybrid assays and immunoblots. Expression of intracellular ECFP/AII similarly activated CREB. However, intracellular and extracellular AII activated CREB through different phosphorylation pathways. Exogenous AII treatment of A10 cells activated p38MAPK and ERK1/2 phosphorylation as determined by Western blot analyses and one-hybrid assays. The p38MAPK inhibitor, SB203580, and the ERK kinase inhibitor, PD98059 each partially inhibited exogenous AII-conferred CREB activation confirming that p38MAPK and ERK1/2 mediate CREB phosphorylation in this system. In contrast, expression of ECFP/AII (intracellular AII) in A10 VSMCs activated p38MAPK but not ERK1/2; inhibition of p38MAPK by SB203580 inhibited intracellular AII-induced CREB phosphorylation. In summary, extracellular AII stimulates at least one pathway common to intracellular AII. This common pathway, in the case of exogenous AII, likely reflects intracellular signaling following internalization of receptor-ligand complex. Extracellular AII also stimulates a unique pathway, apparently reflecting interaction with plasma membrane-associated AT(1)R.

Animals↗
PubMed2006

An intracrine view of angiogenesis.

Angiogenesis, the generation of new blood vessels from pre-existing vessels, is an integral component of wound healing, responses to inflammation and other physiologic processes. It is also an essential part of tumor growth; in the absence of new vessel formation, tumors cannot expand beyond a small volume. Although much is known about angiogenesis and its regulation, there is no overall theory that describes or explains this process. It is here suggested that the intracrine hypothesis, which ascribes to certain extracellular signaling peptides (whether hormones, growth factors, DNA-binding proteins or enzymes) a role in both intracellular biology and extracellular signaling, can contribute to a more general understanding of angiogenesis. Intracrine factors participate in angiogenesis in the following ways: (1) they can act within the cells that synthesized them (type I intracrine action), (2) they can be secreted and then taken up by their cell of synthesis to act intracellularly (type II intracrine action ), or (3) they can be secreted and internalized by a distant target cell (type III intracrine action). The parallels between the intracrine growth factor mechanisms cancer cells employ in stimulating their own growth and the mechanisms operative in endothelial cell proliferation during angiogenesis ("intracrine reciprocity") are discussed. Collectively, these explorations lead to testable hypotheses regarding the regulation of normal and pathological angiogenesis, and point to similarities between tumor-induced angiogenesis and tissue differentiation.

Animals↗
PubMed2005

The intracrine hypothesis: an update.

The intracellular actions of peptide hormones, growth factors, as well as of extracellular-signaling enzymes and DNA-binding proteins, either within target cells or within their cells of synthesis has been called intracrine action. Although these intracrine moieties are structurally diverse, they share certain characteristics of synthesis and function. This has given rise to the development of a theory of intracrine action which permits testable predictions to be made regarding the functioning of these peptides/proteins. Here the intracrine hypothesis is briefly described and then recent experimental findings which bear on predictions made earlier on the basis of the theory are discussed. These findings provide new support for the intracrine hypothesis.

Animals↗
PubMed2005

Reliability of a medication adherence measure in an outpatient setting.

BACKGROUND: Reliable approaches for measuring antihypertensive medication compliance in the outpatient setting are not readily available. The objective of the current study was to determine the reliability of the Hill-Bone Compliance Scale among elderly hypertensive patients. METHODS: We conducted a cross-sectional survey of community-dwelling patients attending the hypertension section of the Internal Medicine Clinic in a large multispecialty group practice. Participants (n=239) completed a self-administered questionnaire consisting of demographic questions and the Hill-Bone Compliance to High Blood Pressure Therapy Scale, which includes a nine-item medication compliance subscale. RESULTS: The mean age of respondents was 69 years; 51% of patients were men, 73% were white, 86% had at least a high school education, and 61% were married. The Cronbach alpha was 0.68 for the medication compliance subscale. All nine items of the medication compliance subscale maintained higher correlations with their own subscale total than with the salt intake and appointment keeping subscale totals. After adjusting for other demographic variables, the odds ratio (95% confidence interval) of perfect medication compliance as reported on the medication compliance subscale was 1.71 (0.95-3.07) for participants 65 years of age and older versus those younger than 65 years of age, 2.53 (1.37-4.66) for whites versus nonwhites, 1.27 (0.73-2.20) for males versus females, 1.30 (0.73-2.29) for married versus unmarried participants, and 1.63 (0.74-3.62) for those with at least a high school education versus those with less education. CONCLUSION: The medication compliance subscale of the Hill-Bone Compliance Scale appears reliable and may be a useful tool for detecting noncompliant patients in outpatient settings.

Aged↗
PubMed2004

The inhibition of tumor growth by triplex-forming oligonucleotides.

We have previously shown that oligonucleotides designed to bind in triplex fashion to a specific p53 binding site homology inhibit the proliferation of colon cancer cells in vitro. The present study was designed to extend these observations in an in vivo model. HCT 116 human colon carcinoma cells were injected subcutaneously into Ncr nude mice and tumors formed at one to two weeks. Tumors were injected daily for 14 days with either triplex forming oligonucleotide (Hoog 1), a scrambled Hoog 1 oligonucleotide (Hoog3) as control, or vehicle. Tumor size was measured twice weekly. Active triplex forming oligonucleotide (Hoog1) reduced tumor size in comparison to either control oligonucleotide (Hoog3) or vehicle. Tumor sizes in the three groups were significantly different (P < 0.001). Student Newman Keuls test shows statistically significant differences between the experimental group and each of the control and vehicle groups (P < 0.05). A triplex forming oligonucleotide directed at a p53 consensus binding site reduces tumor growth suggesting a novel method of tumor inhibition.

Animals↗
PubMed2004

A proposal regarding the biology of memory: participation of intracrine peptide networks.

Although progress has recently been made in defining its neurobiological underpinnings, memory remains an incompletely understood process. Here it is proposed that intracrine peptide action plays a role in memory and therefore in consciousness. Intracrine function is the intracellular action of a peptide either in its cell of synthesis or in target cells following internalization. Intracrine functionality has been associated with growth factors, transcription factors, and enzymes. Some intracrines have been shown to modify their own synthesis, the synthesis of components of their signaling cascades, and/or the synthesis of other intracrines, making it likely that these factors participate in peptinergic regulatory loops/networks. Moreover, some intracrines can affect cellular ion flows over short time periods, while stimulating protein synthesis and growth over longer periods. Intracrines have been reported in the brain (for example, angiotensin, brain derived neurotrophic factor, reelin, and fibroblast growth factor 2) and it is here suggested that intracrine expression/action occurs commonly in the nervous system and, in particular, in the cerebral cortex. This intracrine action leads to the formation of long-lived intra- and intercellular intracrine feed back loops. The intracrine networks thus formed participate in producing both short and long term synaptic potentiation; the former effect is primarily related to intracrine effects on ion flows, the latter to intracrine directed protein synthesis. In addition, intracrine networks provide a previously unappreciated dynamic substrate for the integration of information and ultimately for the formation of the associations that underlay consciousness.

Animals↗
PubMed2004

Tissue renin angiotensin systems.

The RAAS is a powerful regulator of vascular tone and intravascular volume and of tissue architecture and a variety of other functions. The recent appreciation of the immunoregulatory role of angiotensin II and its possible involvement in the genesis of atherosclerosis and in plaque rupture all speak to the wide-ranging physiologic and pathophysiologic activities of the peptide. So do its actions in fat cell differentiation and in neuromodulation. The system exists in the circulation, and RAASs, whole or partial, exist in many tissues. These systems are regulated at many levels ranging from the synthesis of renin to the dimerization of angiotensin receptors. Regulation occurs in multiple tissues and, as a result, tissue concentrations of angiotensin II and the concentration of other RAS components and their active metabolites can vary independently of the circulating system in these tissues. An RAS seems also to function within certain cells. Therapeutic interventions involving ACEIs and ARBs seem likely to provide benefit at least in part through the interruption of local systems. It is to be expected that with enhanced understanding of the biology of the multiple RASs, new suggestions for therapeutic interventions will be forthcoming.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed2004

Mechanisms of disease: local renin-angiotensin-aldosterone systems and the pathogenesis and treatment of cardiovascular disease.

Accumulating evidence has made it clear that not only does the renin-angiotensin-aldosterone system (RAAS) exist in the circulation where it is driven by renal renin, but it is also active in many tissues-and likely within cells as well. These systems might not be completely independent of each other, but rather interact. These local RAASs affect tissue and cellular angiotensin II concentrations and appear to be associated with clinically relevant physiologic and pathophysiologic actions in the cardiovascular system and elsewhere. Evidence in support of this possibility is reviewed here. In addition, direct (pro)renin action after binding to its specific receptor, the existence of renin transcripts, which apparently encode an intracellular renin, the discovery of an angiotensin-converting-enzyme homologue (ACE2), which leads to enhanced generation of angiotensin-(1-7) and the newly appreciated role of angiotensin-receptor dimerization in the regulation of angiotensin activity, all point to the conclusion that the RAASs are complexly regulated, multifunctional systems with important roles both within and outside the cardiovascular system.

Angiotensin II↗
PubMed2003

Intracellular renin and the nature of intracrine enzymes.

Recently, the binding of renin and prorenin to cellular receptors with the subsequent generation of second messengers and the production of physiological effects has been demonstrated. In addition, the internalization of prorenin by target cells has been associated with increased cellular synthesis of angiotensin and cardiac pathology. Also, a renin transcript lacking the sequences encoding a secretory signal has been reported, and this transcript appears to produce a renin that acts in the cell that synthesized it. Some years ago, we coined the term intracrine for a peptide hormone or factor that acts in the intracellular space either after internalization or retention in its cell of synthesis. Thus defined, a wide variety of peptides display intracrine functionality, including hormones, growth factors, transcription factors, and enzymes. For example, considerable evidence indicates that angiotensin II is an intracrine. Also, general principles of intracrine functionality have been developed. Thus, recent evidence demonstrates that the prorenin/renin molecule is an intracrine enzyme. Here, the actions of intracrine enzymes (angiogenin, phosphoglucose isomerase, phospholipase A2, granzyme A and B, thioredoxin, platelet-derived endothelial growth factor, and serine protease inhibitors) are reviewed. The relation of prorenin/renin to other intracrine enzymes, and to intracrines in general, is discussed.

Animals↗
PubMed2003

The intracrine hypothesis and intracellular peptide hormone action.

There is evidence that many peptide growth factors and hormones act in the intracellular space after either internalization or retention in their cells of synthesis. These factors, commonly called intracrines, are structurally diverse while sharing some common functional features. Reports of intracellular peptide hormone binding and action are reviewed here. Also, this laboratory has made proposals regarding the origin and actions of intracrines and these areas are further explored. Intracrine interactions and the relationship of intracrines to transcription factors are discussed. The intracellular/intracrine renin-angiotensin system (iRAS) is reviewed to illustrate the intracrine analogue of a well-established physiological system. The role of intracrine action in metazoan development is also considered.

Animals↗
PubMed2003

Accuracy of telemedicine in detecting uncontrolled hypertension and its impact on patient management.

This study was aimed at assessing the diagnostic accuracy of telemedicine among hypertensive patients. This was a cross-sectional analysis of patients attending a hypertension clinic over a year-long study. Patients were seen both by telemedicine and in-person on the same day with order of the encounters randomly determined. A telemedicine system, which utilized phone lines, was employed. For each type of encounter, whether telemedicine (TM) or in-person (IP), clinical data on blood pressure (BP) control as well as physician ordering patterns were collected. Receiver Operator Characteristic (ROC) curves were used to assess the validity of TM as compared to IP in the assessment of uncontrolled hypertension. Sixty-two patients participated resulting in 107-paired visits over the year-long study period. The mean age of the 62 participants was 67.1 +/- 11.4 years; 56.6% were men. ROC curves for detecting elevated mean blood pressure provided an area under the curve (auc) of 0.87 (95% CI, 0.80-0.95). ROC curves for the detection of uncontrolled systolic hypertension provided an auc of 0.86 (95% CI, 0.78-0.93). Telemedicine-determined BP differed slightly, but statistically significant (p < 0.05), from IP assessments. Meanwhile, there was no difference in ordering diagnostic tests or therapeutics detectable between the two encounter types. Telemedicine proved to be a valid means for detecting uncontrolled BP among hypertensive patients.

Aged↗
PubMed2002

Toward a theory of intracrine hormone action.

A growing body of evidence indicates that in some cases, peptide hormones can function in the intracellular space. These findings are reviewed. In addition, this laboratory has made proposals regarding the origin, nature and function of intracrines--that is, intracellularly acting peptide hormones that also function in an autocrine, paracrine or endocrine manner. Here, these hypotheses are developed, and potential implications/applications of this point of view are discussed. Possible implications for cellular differentiation, cellular memory and hormonal responsiveness, as well as for the assumption of novel functions by intracellular regulatory proteins are discussed.

Angiotensins↗
PubMed2002

Intracellular angiotensin II increases the long isoform of PDGF mRNA in rat hepatoma cells.

Our recent published studies suggest that angiotensin II (AII), generated and retained intracellularly, enhances growth of H4-II-E-C3 rat hepatoma cells, an average of 33%. Proliferation conferred by introduction of a plasmid [ Ang(-S)Exp/pSVL ] encoding a signal sequence-depleted angiotensinogen [Ang(-S)Exp] into these cells (which we have shown possess ACE and renin mRNAs) is mediated, at least in part, by enhanced PDGF-A chain mRNA production and protein secretion. The mitogenic effect is inhibited by losartan suggesting that it involves AII interaction with an AT(1)-like receptor. Introduction of anti-AII antibodies into the medium of these transfected cells has no effect upon growth of the cells, suggesting that AII is retained by the cells and that intracellular AII is growth stimulatory. In the present study, we sought to further characterize the intracellular localization and mode of action of Ang(-S)Exp. Consistent with our expectations, we now show that a fusion product of Ang(-S)Exp with green fluorescent protein [Ang(-S)Exp/EGFP], generated from an expression plasmid, is abundant and primarily cytoplasmic. Wild-type angiotensinogen/EGFP, in contrast, is only detectable following a cold-block (which acts to enhance folding-kinetics and slow secretion) and is largely restricted to the secretory pathway. We further show, using semi-quantitative RT/PCR that the long isoform of PDGF mRNA is elevated in Ang(-S)Exp transfected cells and in AII-treated naive cells but not in losartan-treated Ang(-S)Exp transfected cells. We identify C-terminal amidation recognition sites within the long-form protein (that are not present in the short-form) and show that these cells possess PAM (amidating enzyme precursor) and carboxypeptidase E mRNAs (the corresponding proteins of which are sufficient for amidation). Inhibitors of amidation inhibit growth of naive and Ang(-S)Cntr/ pSVL -transfected cells (2.6-fold for phenylbutenoic acid and 3.5-fold for disulfiram treatment) but more profoundly inhibit growth of Ang(-S)Exp/pSVL -transfected cells (6.7-fold for phenylbutenoic acid and 13-fold for disulfiram). In conclusion, these data confirm that signal sequence-depleted Ang(-S)Exp is retained within cells and is largely cytoplasmic. Because C-terminal amidation is absolutely required for full biological potency of a number of peptide hormones (including oxytocin, gastrin and calcitonin), we postulate that growth effects of both intracellular AII and exogenous AII can be conferred by PDGF long-form, possibly through an amidation-dependent mechanism.

Amides↗
PubMed2002

The origins of intracrine hormone action.

A growing number of peptide hormones and growth factors have been shown to operate in the intracellular space after either internalization or retention in their cells of synthesis. These factors, called intracrines, in many cases are expressed as multiple isoforms, traffic to nucleus or nucleolus, and regulate gene transcription. Some intracrines are angiogenic. It is here argued that intracrine action is the modern analogue of a biologically ancient mechanism for regulating message translation and ribosome synthesis. The implications of this view for research and therapeutics are discussed.

Animals↗
PubMedDate not supplied

Application of the combined quality improvement ratio in the evaluation of a quality improvement activity in a managed care organization.

Assessing the impact of clinically relevant quality improvement activities (QIA) is important to managed care organizations; yet, turnover in enrollment can reduce the data available for analyses, thus decreasing the chance that a difference post-QIA will be detected. The Combined Quality Improvement Ratio (CQuIR) uses matching of patients into pairs pre- and post-QIA to systematically and validly increase the data included in the analysis for evaluation of the QIA. Using a paired cohort study design, 456 pairs of patients with diabetes were identified using the Health Plan Employer Data Information Set (HEDIS) specifications. Patients having retinal examinations were identified pre- and post-QIA. The change in retinal examination rates was analyzed and results compared using repeated pairs (RP), matched pairs (MP), and combined pairs (CP). The CQuIR methodology (which uses CP = RP + MP) resulted in an increase in sample size (n = 456 [CP] versus n = 156 [RP] and n = 300 [MP]) and consequently an increase in power (0.92 [CP] versus 0.38 [RP] and 0.82 [MP]) and a decrease in the confidence interval range (0.97 [CP] versus 2.06 [RP] and 1.14 [MP]). The CQuIR uses a statistically valid approach to increase the data available for the evaluation of QIAs.

Adult↗