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Biomedical subjects

R A Johnson

Publications and source records attributed to R A Johnson.

At least 55 records · Page 3Linked to original sources
PubMed2000

Epstein-Barr virus immediate-early proteins BZLF1 and BRLF1 activate the ATF2 transcription factor by increasing the levels of phosphorylated p38 and c-Jun N-terminal kinases.

Expression of either Epstein-Barr virus (EBV) immediate-early protein BZLF1 (Z) or BRLF1 (R) is sufficient to convert EBV infection from the latent to lytic form. Disruption of viral latency requires transcriptional activation of the Z and R promoters. The Z and R proteins are transcriptional activators, and each immediate-early protein activates expression of the other immediate-early protein. Z activates the R promoter through a direct binding mechanism. However, R does not bind directly to the Z promoter. In this study, we demonstrate that the ZII element (a cyclic AMP response element site) in the Z promoter is required for efficient activation by R. The ZII element has been shown to be important for induction of lytic EBV infection by tetradecanoyl phorbol acetate and surface immunoglobulin cross-linking and is activated by Z through an indirect mechanism. We demonstrate that both R and Z activate the cellular stress mitogen-activated protein (MAP) kinases, p38 and JNK, resulting in phosphorylation (and activation) of the cellular transcription factor ATF2. Furthermore, we show that the ability of R to induce lytic EBV infection in latently infected cells is significantly reduced by inhibition of either the p38 kinase or JNK pathways. In contrast, inhibition of stress MAP kinase pathways does not impair the ability of Z expression vectors to disrupt viral latency, presumably because expression of Z under the control of a strong heterologous promoter bypasses the need to activate Z transcription. Thus, both R and Z can activate the Z promoter indirectly by inducing ATF2 phosphorylation, and this activity appears to be important for R-induced disruption of viral latency.

Activating Transcription Factor 1↗
PubMed2000

Are pesticides really endocrine disruptors?

In recent years, a considerable amount of basic laboratory investigation has been targeted toward understanding the relationship between environmental contaminants and cellular endocrine function. A number of environmental contaminants are now known to alter endocrine physiology without acting as classic toxicants. Hence, a new field of inquiry has emerged within the discipline of environmental toxicology, the study of endocrine disrupting chemicals (EDCs). Pesticides represent one of the better studied groups of EDCs. As the tools of molecular biology become increasingly sophisticated, our ability to understand the endocrine effects of these compounds continues to broaden at a remarkable rate. Still, clinical data linking them to derangements in human endocrine function have been quite limited. While the scientific community awaits further epidemiological assessment of the impact of pesticides on public health, responsible land management practices should be employed in an effort to reduce the burden of these chemicals ultimately reaching our water supply.

Endocrine System↗
PubMed2000

The optimum time to employ telephotoscreening to detect retinopathy of prematurity.

PURPOSE: Labor-intensive screening of infants in the neonatal intensive care units is the only way to detect retinopathy of prematurity (ROP). Our purpose is to determine if RetCam 120 photos, acquired by a neonatal nurse, can be used to screen for ROP by performing 2 screening examinations, at 32 to 34 weeks (exam 1) and at 38 to 40 weeks (exam 2) post-conceptional age. METHODS: RetCam examinations are performed by a nurse on infants at exam 1 and exam 2 intervals. At the same time, an examination is performed by an experienced ophthalmologist. Masked readers evaluate the photos for ROP and determine if each eye will progress to prethreshold or threshold disease. The data are compared to the clinical course of the eyes. RESULTS: Forty-six eyes were photographed at exam 1 and 50 eyes at exam 2 from July 1, 1999, to December 15, 1999. Sensitivity and specificity of detecting ROP were 76% and 100% for exam 2 and 46% and 100% for exam 1. Sensitivity and specificity of predicting prethreshold disease were 64% and 97% for exam 2 and 33% and 100% for exam 1. Sensitivity and specificity of predicting threshold were both 100% at exam 2 and 0% (one photo in category) and 95% at exam 1. CONCLUSION: A potential reason for low sensitivity is technical limitations of the Retcam, such as the difficulty in capturing peripheral retina in small eyes and the need for a better lid speculum.

Disease Progression↗
PubMed2000

Adolescent cigarette smoking in U.S. racial/ethnic subgroups: findings from the National Education Longitudinal Study.

Using nationally representative data for 16,454 8th graders and 13,840 10th graders, we explore racial/ethnic differences in "daily cigarette initiation," beginning to smoke on a daily basis between baseline interviews and reinterviews conducted two years later. In both samples, the initiation rate among whites is more than double the rate among blacks and higher than rates among Asian/Pacific Islanders and Hispanics. Risk factors at the individual, family, and peer-group levels of analysis do not explain most racial/ethnic differences. We develop alternative hypotheses by extending theories of cigarette use to the school level, and we test them using multilevel models: Consistent with social learning theory, cigarette risk among blacks and Hispanics decreases as the percentage of racial/ethnic minority students in the school increases. Consistent with strain theory, cigarette risk increases with the academic competitiveness of the school--especially among females--after controlling for the adolescent's academic performance.

Adolescent↗
PubMed2000

Administrative channeling under the Medicare Act clarified: Illinois Council, section 405(h), and the application of congressional intent.

Mr. Cogan and Mr. Johnson discuss the judicial review provisions of the Medicare Act, codified at 42 U.S.C. section 405(g) and (h), and the Supreme Court cases including Illinois Council that interpret the scope of the "arising under" language. They also examine the history of section 205(h) of the Social Security Act, including the most recent amendment contained in the Deficit Reduction Act of 1984. The relationship between 205(h), DEFRA, and section 405(h) is explored as well as the caselaw addressing and applying section 405(h) in light of Congress' 1984 amendments.

Bankruptcy↗
PubMed1999

Enzymatic synthesis of unlabeled and beta-(32)P-labeled beta-L-2', 3'-dideoxyadenosine-5'-triphosphate as a potent inhibitor of adenylyl cyclases and its use as reversible binding ligand.

beta-L-2',3'-Dideoxyadenosine-5'-triphosphate (beta-L-2', 3'-dd-5'-ATP) was prepared enzymatically from the corresponding monophosphate by the use of adenylate kinase, creatine phosphate, and creatine kinase in a single step. The beta-(32)P-labeled analog was prepared similarly, but in a two step reaction. beta-L-2', 3'-dd-5'-ATP inhibited adenylyl cyclase from rat brain competitively with respect to substrate (5'-ATP.Mn(2+)) and exhibited an IC(50) approximately 24 nM. The labeled ligand was used in the development of a reversible binding assay for adenylyl cyclases. Binding of beta-L-2',3'-dd-[beta-(32)P]5'-ATP was saturable with increasing concentrations of ligand and increased in proportion to membrane protein, and was enhanced by Mn(2+) to a greater extent than by Mg(2+). Binding was displaced with adenine nucleotides known to be either competitive or noncompetitive inhibitors but not by agents known not to act on the cyclase, or by 3-isobutyl-1-methylxanthine, creatine phosphate, or creatine kinase. Binding was rapid, with a half-time for the on-rate <1.8 min and for the off-rate <0.8 min. The potency and mechanism of the inhibition of this ligand and the pattern of agents that displace binding suggest an interaction with adenylyl cyclase per se and to a configuration of the enzyme consistent with an interaction at the catalytic active site. The data suggest that this is a pretransition state inhibitor and contrasts with the equipotent 2',5'-dd-3'ATP, a post-transition state noncompetitive inhibitor.

Adenosine Triphosphate↗
PubMed1999

Inhibition of adenylyl cyclase by acyclic nucleoside phosphonate antiviral agents.

Acyclic derivatives of adenine, known as highly effective nucleotide analogs with broad spectrum antiviral activity, were evaluated for potential cross-reactivity with adenylyl cyclases, a family of membrane-bound enzymes that share putative topologies at their catalytic sites with oligonucleotide polymerases and reverse transcriptases. A series of derivatives of 9-(2-phosphonylmethoxyethyl)adenine (PMEA) inhibited a preparation of adenylyl cyclase derived from rat brain with IC(50) values that ranged from 66 microM (PMEA) to 175 nM for its diphosphate derivative (PMEApp) and mimics of it. PMEApp mimics included PMEAp(NH)p, PMEAp(CH(2))p, PMEAp(CX(2))p (X = fluorine, chlorine, or bromine), PMEAp(CHX)pp, and PMEAp(C(OH)CH(3)pp. The data suggest that inhibition of adenylyl cyclases may contribute to the therapeutic action of some of these or similar compounds or constitute part of their side effects in therapeutic settings.

Adenine↗
PubMed1999

Covalent labeling of adenylyl cyclase cytosolic domains with gamma-methylimidazole-2',5'-dideoxy-[gamma-(32)P]3'-ATP and the mechanism for P-site-mediated inhibition.

A truncated first cytosolic domain of type V adenylyl cyclase (VC(1)) and a truncated second cytosolic domain of type II adenylyl cyclase (IIC(2)) were used alone and in the readily reversible complex (VC(1).IIC(2)) to evaluate interactions with each other and with reversible and irreversible P-site ligands. Enzyme activity was used to assess formation and dissolution of VC(1).IIC(2). The data suggest that binding of 2',5'-dideoxy-3'-ATP to VC(1) and IIC(2) prevented formation of VC(1).IIC(2) and that 2',5'-dideoxy-3'-ATP dissociation occurred slowly. To enable configuration specific cross-linking to the catalytic site, 2',5'-dideoxyadenosine 3'-[gamma-(1-methylimidazole)-triphosphate] (gamma-MetIm-2', 5'-dd-3'-ATP) and 2',5'-dd-adenosine 3'-(gamma-azidoanilido)-triphosphate (gamma-azidoanilido-2', 5'-dd-3'-ATP) were synthesized, the former also as its gamma-(32)P-labeled analog. gamma-Azidoanilido-2',5'-dd-3'-ATP exhibited an inhibitory potency comparable with that of 2', 5'-dd-3'-ATP. gamma-MetIm-2',5'-dd-[gamma-(32)P]3'-ATP labeled the individual VC(1) and IIC(2) domains comparably and covalently to approximately 20% within 1 h. Formation of VC(1).IIC(2) resulted in reduced labeling of VC(1) but enhanced labeling of IIC(2). The data imply that formation of the catalytically active VC(1).IIC(2) complex affects the interaction of each domain with the 2', 5'-dd-3'-ATP, the binding of which also affects the interaction between the two cytosolic domains, leading to a pseudo-irreversible inhibition.

Adenosine Triphosphate↗
PubMed1999

Two-metal-Ion catalysis in adenylyl cyclase.

Adenylyl cyclase (AC) converts adenosine triphosphate (ATP) to cyclic adenosine monophosphate, a ubiquitous second messenger that regulates many cellular functions. Recent structural studies have revealed much about the structure and function of mammalian AC but have not fully defined its active site or catalytic mechanism. Four crystal structures were determined of the catalytic domains of AC in complex with two different ATP analogs and various divalent metal ions. These structures provide a model for the enzyme-substrate complex and conclusively demonstrate that two metal ions bind in the active site. The similarity of the active site of AC to those of DNA polymerases suggests that the enzymes catalyze phosphoryl transfer by the same two-metal-ion mechanism and likely have evolved from a common ancestor.

Adenosine Triphosphate↗
PubMed1999

Role of carbon monoxide in L-glutamate-induced cardiovascular responses in nucleus tractus solitarius of conscious rats.

Heme oxygenase degrades heme to form carbon monoxide. It has been reported that heme oxygenase-derived carbon monoxide may interact with L-glutamate (L-Glu) receptors in the nucleus tractus solitarius (NTS). Integrative studies suggest that heme oxygenase inhibitors raise blood pressure, in part, by inhibiting carbon monoxide formation in the NTS. The currents studies were designed to determine if heme oxygenase inhibitors affect the cardiovascular actions of L-Glu in the NTS. Accordingly, MAP and HR responses to unilateral microinjections of L-Glu (5 nmol/100 nl) into the NTS were measured before and after ipsilateral microinjections of zinc deuteroporphyrin 2,4-bis glycol (ZnDPBG, 4.5 nmol/100 nl) or chromium mesoporphyrin (CrMP, 1.5 nmol/100 nl) in awake rats chronically instrumented with NTS guide cannulaes and arterial catheters. With respect to non-treatment (+36+/-5 mmHg, -107 bpm, n=10), ZnDPBG pre-treatment attenuated the pressor and bradycardic responses to L-Glu (+7+/-3 mmHg, -10+/-6 bpm, P<0.05). CrMP similarly attenuated cardiovascular responses to L-Glu (+47+/-3 mmHg, -68+/-8 bpm vs. +20+/-5 mmHg, -40+/-9 bpm; before vs. after, n=10, P<0.05). Matched series yielded no vehicle- or time-related effects. Our findings suggest that a heme oxygenase product, such as carbon monoxide, may affect NTS glutamatergic neurotransmission to participate in cardiovascular control.

Analysis of Variance↗
PubMed1999

Skeletal muscle and small-conductance calcium-activated potassium channels.

Skeletal muscle becomes hyperexcitable following denervation and when cultured in the absence of nerve cells. In these circumstances, the bee venom peptide toxin apamin, a blocker of small-conductance calcium-activated potassium (SK) channels, dramatically reduces the hyperexcitability. In this report, we show that SK3 channels are expressed in denervated skeletal muscle and in L6 cells. Action potentials evoked from normal innervated rat skeletal muscle did not exhibit an afterhyperpolarization, indicating a lack of SK channel activity; very low levels of apamin binding sites, SK3 protein, or SK3 mRNA were present. However, denervation resulted in apamin-sensitive afterhyperpolarizations and increased apamin binding sites, SK3 protein, and SK3 mRNA. Cultured rat L6 myoblasts and differentiated L6 myotubes contained similar levels of SK3 mRNA, although apamin-sensitive SK currents and apamin binding sites were detected only following myotube differentiation. Therefore, different molecular mechanisms govern SK3 expression levels in denervated muscle compared with muscle cells differentiated in culture.

Animals↗
PubMed1999

Inhibitory effect of 4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)1H - imidazole on HCMV DNA replication and permissive infection.

We found that Human Cytomegalovirus (HCMV) infection of human fibroblasts resulted in a dramatic increase in p38 mitogen-activated protein kinase (MAPK) phosphorylation. Recently, drug mediated inhibition of p38 has been demonstrated to exhibit anti-viral activity against HIV (Shapiro, L., Heidenreich, K.A., Meintzer, M.D. and Dinarello, C.A., 1998. Role of p38 mitogen-activated protein kinase in HIV type 1 production in vitro. Proc. Natl. Acad. Sci. USA. 95, 7422-7426). Therefore, we examined the effect of a specific p38 kinase inhibitor on HCMV infection. Inhibiting p38 activity in HCMV infected cells by treating cells with 4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)1H-imidazole; (FHPI), a p38 inhibitor drug, prevented permissive HCMV infection as measured by plaque assay. In the presence of FHPI, HCMV immediate early gene expression was slightly lower at early times of infection, but there was no inhibition of expression of the early gene UL-84, an HCMV protein essential for viral replication. However, FHPI inhibited HCMV DNA replication and late gene expression. The inhibitory effect of FHPI was reversible, as demonstrated by the induction of HCMV replication upon withdrawal of FHPI. Our data describes FHPI as a novel anti-HCMV compound that inhibits synthesis/activation of cellular and/or viral factors required for initiation of HCMV DNA replication.

Antiviral Agents↗
PubMed1999

Prevalence of skin disease in a cohort of shelter-based homeless men.

BACKGROUND: Cutaneous disease is a frequent cause of morbidity in homeless people. A variety of infectious and noninfectious conditions have been described in this population that are exacerbated by malnutrition, exposure to hazardous environment, psychiatric disease, physical injuries, and limited access to health care. OBJECTIVE: Our purpose was to study the prevalence of skin disease in a selected group of the homeless population. METHODS: We evaluated skin diseases in a cohort of 142 homeless men who were staying overnight in a major Boston homeless shelter for men over a period of 3 months (March-June 1992). The subjects participated in the study by volunteering for a free skin examination. Completion of a detailed questionnaire on age, race, duration of homelessness, and general medical status was followed by a thorough skin examination performed by dermatologists. RESULTS: The most prevalent skin diseases in the examined population were tinea pedis (38%), pitted keratolysis of the feet (20.4%), traumatic injuries (19.7%), toenail onychomycosis (15.4%), acne vulgaris (18.3%), and seborrheic dermatitis (13.3%). CONCLUSION: To our surprise the majority of patients had relatively normal findings from skin examinations with only a few pathologic findings, most often related to poor foot hygiene and long-term exposure to moisture. We attribute the relatively good condition of our cohort to the excellent care provided by the shelter with regard to clothing, cleaning facilities, and medical care. Our findings may represent the prevalence of skin diseases in shelter-based homeless but not in street homeless persons who are likely to have much poorer hygiene and perhaps more skin disease.

Adult↗