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S A Kane

Publications and source records attributed to S A Kane.

At least 19 recordsLinked to original sources
PubMed2001

Characterisation of the effects of a non-peptide CGRP receptor antagonist in SK-N-MC cells and isolated human cerebral arteries.

The cerebral circulation is innervated by calcitonin gene-related peptide (CGRP) containing fibers originating in the trigeminal ganglion. During a migraine attack, there is a release of CGRP in conjunction with the head pain, and triptan administration abolishes both the CGRP release and the pain at the same time. In the search for a novel treatment of migraine, a non-peptide CGRP antagonist has long been sought. Here, we present data on a human cell line and human and guinea-pig isolated cranial arteries for such an antagonist, Compound 1 (4-(2-Oxo-2,3-dihydro-benzoimidazol-1-yl)-piperidine-1-carboxylic acid [1-(3,5-dibromo-4-hydroxy-benzyl)-2-oxo-2-(4-phenyl-piperazin-1-yl)-ethyl]-amide). On SK-N-MC cell membranes, radiolabelled CGRP binding was displaced by both CGRP-(8-37) and Compound 1, yielding pK(i) values of 8.9 and 7.8, respectively. Functional studies with SK-N-MC cells showed that CGRP-induced cAMP production was antagonised by both CGRP-(8-37) and Compound 1 with pA(2) values of 7.8 and 7.7, respectively. Isolated human and guinea pig cerebral arteries were studied with a sensitive myograph technique. CGRP induced a concentration-dependent relaxation in human cerebral arteries which was antagonized by both CGRP-(8-37) and Compound 1 in a competitive manner. In guinea pig basilar arteries, CGRP-(8-37) antagonised the CGRP-induced relaxation while Compound 1 had a weak blocking effect. The clinical studies of non-peptide CGRP antagonists are awaited with great interest.

Animals↗
PubMed2001

Cloning, characterization and central nervous system distribution of receptor activity modifying proteins in the rat.

Calcitonin gene-related peptide (CGRP), adrenomedullin (ADM), amylin and calcitonin (CT) are structurally and functionally related neuropeptides. It has recently been shown that the molecular pharmacology of CGRP and ADM is determined by coexpression of one of three receptor activity-modifying proteins (RAMPs) with calcitonin receptor-like receptor (CRLR). Furthermore, RAMP proteins have also been shown to govern the pharmacology of the calcitonin receptor, which in association with RAMP1 or RAMP3, binds amylin with high affinity. In this study, we have cloned the rat RAMP family and characterized the pharmacology of rat CGRP and ADM receptors. Rat RAMP1, RAMP2 and RAMP3 shared 72%, 69% and 85% homology with their respective human homologues. As expected CRLR-RAMP1 coexpression conferred sensitivity to CGRP, whilst association of RAMP2 or RAMP3 with CRLR conferred high affinity ADM binding. Using specific oligonucleotides we have determined the expression of RAMP1, RAMP2 and RAMP3 mRNAs in the rat central nervous system by in situ hybridization. The localization of RAMP mRNAs was heterogeneous. RAMP1 mRNA was predominantly expressed in cortex, caudate putamen and olfactory tubercles; RAMP2 mRNA was most abundant in hypothalamus; and RAMP3 was restrictively expressed in thalamic nuclei. Interestingly, in specific brain areas only a single RAMP mRNA was often detected, suggesting mutual exclusivity in expression. These data allow predictions to be made of where each RAMP protein may heterodimerize with its partner G-protein-coupled receptor(s) at the cellular level and consequently advance current understanding of cellular sites of action of CGRP, ADM, amylin and CT. Furthermore, these localization data suggest that the RAMP family may associate and modify the behaviour of other, as yet unidentified neurotransmitter receptors.

Adrenomedullin↗
PubMed2001

Functional role of alpha-calcitonin gene-related peptide in the regulation of the cardiovascular system.

It remains unknown whether the extent of vasoactive response to exogenous calcitonin gene-related peptide (CGRP) varies among different regional vascular beds. It is also unclear whether endogenous CGRP plays a functional role in regulating basal vascular activity. To address these two issues, experiments were conducted in 27 anesthetized rats instrumented with a carotid flow probe and catheters in a jugular vein, left ventricle (LV), and femoral artery, and in 6 conscious dogs, chronically instrumented with LV pressure gauge, aortic and atrial catheters, and ascending aortic, coronary, carotid, and renal flow probes. In both species, administration of human alpha-CGRP (0.1-0.5 microg/kg, i.v.) induced a dose-dependent peripheral vasodilation that was completely abolished by pretreatment with alpha-CGRP[8-37] (30 microg/kg/min, i.v.), a competitive antagonist of CGRP receptors. Regional blood flow measured by the radioactive microsphere technique in rats showed that the alpha-CGRP (0.3 microg/kg, i.v.)-induced increase in blood flow was greater (p < 0.05) in the heart (+53 +/- 16%) than in the brain (+14 +/- 6%). In the presence of beta-adrenergic receptor blockade with propranolol, however, the increases in blood flow in these two vascular beds were identical. In conscious dogs, alpha-CGRP (0.3 microg/kg, i.v.) produced similar increases in coronary (+24 +/- 6%), carotid (+26 +/- 3%), and renal (+26 +/- 6%) blood flow, which were different from the patterns induced by other vasodilators; at an equivalent level of reduction in mean arterial pressure and total peripheral resistance, alpha-CGRP increased coronary and carotid blood flow significantly less (p < 0.05) than adenosine or nitroprusside. Unlike alpha-CGRP, adenosine and nitroprusside, as expected, induced pronounced differential blood flow changes in these vascular beds. Neither systemic hemodynamics nor regional blood flow distribution was altered by the administration of a pharmacological blocking dose of alpha-CGRP[8-37] in the two species. Thus, we conclude that endogenous alpha-CGRP does not play an important role in cardiovascular regulation under normal, resting conditions, although exogenous alpha-CGRP induces a marked, comparable vasorelaxation in different regional vascular beds.

Adenosine↗
PubMed2000

Development of a binding assay for p53/HDM2 by using homogeneous time-resolved fluorescence.

The p53 tumor suppressor protein is activated and stabilized in response to DNA damage, resulting in cell cycle arrest or apoptosis. HMD2 is a negative regulator of p53. Binding of p53 by HDM2 traffics p53 from the nucleus to the cytoplasm where it is recognized and targeted for ubiquitin-mediated degradation (D. A. Freedman, L. Wu, and A. J. Levine, 1999, Cell. Mol. Life Sci. 55, 96-107). Several reports have suggested that disruption of this complex in normal cells results in p53 signaling (V. Böttger, A. Böttger, A. Sparks, W.-L. Liu, S. F. Howard, and D. P. Lane, 1997, Curr. Biol. 7, 860-869; C. Wasylyk, R. Salvi, M. Argentini, C. Dureuil, I. Delumeau, J. Abecassis, L. Debussche, and B. Wasylyk, 1999, Oncogene 18, 1921-1934). A homogeneous time-resolved fluorescence (HTRF) assay has been developed to monitor p53/HDM2 binding. This assay employs a site-specific biotinylated p53 protein, a GST-fused HDM2 protein, and two fluorophore-conjugated detection reagents, streptavidin-XL665 and europium cryptate-labeled anti-GST antibody ¿Eu(K)-anti-GST. Binding of p53 to HDM2 brings the fluorophores into close proximity, allowing fluorescence resonance energy transfer to occur. Development of this assay and comparison to a traditional ELISA are described in this report. The HTRF assay was then utilized to assess the effect of serine phosphorylation within the p53 N-terminus on HDM2 binding, and to determine the relative affinity of a p73 peptide for HDM2.

Amino Acid Sequence↗
PubMed1999

Activation of K-p21ras and N-p21ras, but not H-p21ras, is necessary for mitogen-induced human airway smooth-muscle proliferation.

Ras proteins (H-, K-, and N-p21ras) play critical roles in the control of normal and neoplastic cell growth. To date, however, little is known about the role of p21ras in regulating mitogen-induced smooth muscle and, specifically, human airway smooth-muscle (HASM) cell growth. We postulate that p21ras is a critical signaling event regulating mitogen-induced HASM cell proliferation. Growth-arrested, confluent HASM cells were treated for 1 h with 10 ng/ml epidermal growth factor (EGF), 1 U/ml thrombin, or 5 microM bradykinin, then cell lysates were immunoprecipitated using anti-p21ras antibody. Immunoblot analysis using a pan p21ras antibody, which recognizes H-, K-, and N-p21ras, found no significant difference in p21ras expression in HASM after stimulation with either agent, as compared with control. In parallel experiments, we characterized that HASM cells express K- and N-p21ras, but not H-p21ras. Further, there was no difference between the levels of each p21ras isoform after stimulation with any of the agonists. The time course of p21ras activation, however, was markedly different among agonists. EGF rapidly activated p21ras within 30 s and was sustained for up to 30 min. Although thrombin also induced a rapid rise in p21ras activity after 2.5 min, the activation was transient. In contrast, bradykinin, which is nonmitogenic for HASM cells, did not activate p21ras. Using single-cell microinjection, the role of p21ras activation in modulating mitogen-induced HASM DNA synthesis was determined by 5-bromo-2'-deoxyuridine (BrdU) incorporation and anti-BrdU immunofluorescent staining. Thrombin- and EGF-induced DNA synthesis in cells microinjected with Y13-259, a neutralizing p21ras antibody, was significantly inhibited as compared with those microinjected with isotype-matched rat immunoglobulin G(1) or a vehicle control. These data suggest that activation of p21ras appears to be necessary for EGF and thrombin-induced HASM cell proliferation and that activation of K- and N-p21ras, but not H-p21ras, mediates smooth-muscle cell growth.

Antibodies↗
PubMed1997

Magnetic resonance imaging of optic perineuritis.

Optic perineuritis, an uncommon variant of orbital pseudotumor, may be clinically indistinguishable from retrobulbar optic neuritis. Because treatment and prognosis for these two entities are different, early diagnosis is important. We report a case of a 47 year-old woman with clinical findings suggestive of retrobulbar optic neuritis, but whose magnetic resonance images suggested optic perineuritis. A dramatic clinical response to oral corticosteroids was observed. Optic perineuritis should be considered in cases of presumed retrobulbar optic neuritis. MRI may differentiate these two entities in the acute stage, and should be considered before treatment is decided.

Contrast Media↗
PubMed1996

Photoreactivity of platinum(II) in cisplatin-modified DNA affords specific cross-links to HMG domain proteins.

Cisplatin-modified DNA forms specific complexes with proteins that contain the DNA binding motif known as the high-mobility group (HMG) domain. As a tool for investigating the role of these proteins in mediating the cytotoxic effects of cisplatin, a set of cisplatin analogs was prepared in which one of the ammine ligands was replaced with a photoreactive tethered aryl azide ligand. The ability of DNA modified by these platinum complexes to photo-cross-link to HMG1 was investigated. During this study, it was discovered that DNA modified with cisplatin itself can undergo photoinduced cross-linking to HMG1 when irradiated with 300 nm light. The covalent complexes resulting from this latter cross-linking reaction are completely reversed by the addition of sodium cyanide and can be degraded by proteinase K. These results confirm the presence of a protein-DNA cross-link and demonstrate that the platinum atom itself forms the point of attachment. By contrast, DNA modified with transdiamminedichloroplatinum(II), [Pt(dien)Cl]Cl, or [Pt(NH3)3Cl]Cl does not cross-link to HMG1 upon irradiation. The photochemistry was exploited to cross-link a 15-base pair oligonucleotide containing a single, site-specific cis-[Pt(NH3)2{d(GpG)-N7(1),-N7(2)}] intrastrand adduct to domain B of HMG1. Following proteolytic digestion of the resulting covalent complex, the site of attachment to the protein was determined by Edman degradation of the resulting peptide-DNA complex to be a single residue on HMG domain B, Lys-6. The data further suggest that this amino acid binds to platinum at a site made available by photolabilization of a purine ligand. These results afford the first structural information about the interaction of HMG domain proteins with cisplatin-modified DNA.

Amino Acid Sequence↗
PubMed1995

Specific cleavage of a DNA triple helix by FeII.bleomycin.

The specific cleavage of a DNA triple helix by FeII.bleomycin (BLM) is demonstrated. Triplex-specific cleavage was observed on both strands of the 32-base pair (bp) duplex at the duplex-triplex junctions. Strand scission products and alkali labile lesions were both formed. The strongest BLM cleavage site was located at the 5'-duplex-triplex junction, which is also the preferred triplex binding site of intercalating agents [Collier, D. A., Mergny, J.-L., Thuong, N. T., & Hélène, C. (1991) Nucleic Acids Res. 19, 4219-4224]. The preference of BLM for the 5'-junction does not appear to derive from selective intercalative binding at this site. This is supported by the observation that phleomycin, which contains a thiazolinylthiazole moiety rather than a planar bithiazole ring system, exhibited the same selectivity of triplex cleavage as BLM. Cleavage of the triple helix by FeII.BLM was unaffected by concentrations of Mg2+ up to 5 mM, suggesting possible therapeutic applications of this novel DNA target. Molecular-modeling calculations of the triplex region suggested that dramatic variations in minor groove width and depth occur at the duplex-triplex junctions, particularly at the 5'-junction. Moreover, the minor groove at these sites was calculated to be somewhat shallower and wider than the minor groove of B-DNA. These results suggest that the preference of BLM for the duplex-triplex junctions derives from selective recognition of minor groove shape at these sites and thus reflects conformation-selective, rather than sequence-selective, DNA recognition by FeII.BLM.

Antibiotics, Antineoplastic↗
PubMed1994

On the role of the bithiazole moiety in sequence-selective DNA cleavage by Fe.bleomycin.

A recent study of Fe(II).bleomycin-mediated DNA strand scission suggested that the metal binding domain of the drug is also the primary determinant of the observed sequence selectivity of strand scission (Carter, B. J., Murty, V. S., Reddy, K. S., Wang, S.-N., and Hecht, S. M. (1990) J. Biol. Chem. 265, 4193-4196). Although it is well established that the bithiazole moiety+C-terminal substituent of bleomycin are required for DNA binding, the role of the bithiazole in sequence-selective DNA recognition remains unclear. To determine whether the bithiazole moiety exhibits an intrinsic DNA binding selectivity, three synthetic EDTA-conjugated bithiazole derivatives were used to mediate DNA cleavage in the presence of Fe2+ and dithiothreitol. Incubation of these Fe(II).EDTA-bithiazoles in the presence of a 5'-32P end-labeled DNA duplex resulted in strand scission at every position to essentially the same extent. The relative cleavage efficiencies among the bithiazoles were a strong function of their ionic state. These findings imply that the bithiazoles can bind to many sites on the DNA; they support a model of bleomycin-DNA interaction in which the bithiazole moiety+C-terminal substituent are required only for DNA binding, whereas the metal binding domain is responsible for metal ion coordination and oxygen activation as well as being the primary determinant of sequence-selective DNA cleavage.

Binding Sites↗
PubMed1993

Cerebellar nuclei: rapid alternating movement, motor somatotopy, and a mechanism for the control of muscle synergy.

Monkeys were trained to make rapid alternating flexion and extension movements seriatim at each of five body parts--foot (toe grasp and release), shoulder, elbow, wrist, and thumb. "Facial movements"--mouth, tongue, jaw and pharynx--were also made to drink the fruit juice reward. Movement at a given joint could be performed simply by alternate activation of the prime mover muscle groups, and EMG analysis indicated that these primary muscles were active during movement of that joint. No muscle within any one large body part (leg or arm) was strongly active in relation to movement of another body part. Yet, within a body part (arm), synergist muscles were often more active than the primaries during movement of a given joint, only to become less active during movement of the joint at which their action was primary. Neurons in dentate and interpositus discharged in relation to these movements. In the antero-posterior dimension of both dentate and interposed nuclei, there was a significant tendency for neural modulation to be somatotopically arranged according to the preferred movement: hindlimb anteriormost, forelimb in the middle, and head posteriorly. In the medio-lateral dimension, no such localization was seen for the different movements of the upper limb. Like muscle activity, neural discharge modulation usually occurred in strong relation to a number of movements in a single body part (arm), but not to the movements of different body parts (leg, face). Lesion of the middle third of dentate and of a portion of lateral interpositus had little effect upon the movements at the single joints, and thus upon the prime movers. However, the pattern of activity of agonist, antagonist, and synergist muscles was changed. These results are consistent with the view that the dentate controls muscle synergy and movement coordination more than the prime movers per se.

Animals↗
PubMed1990

Synthesis of C-glycosylarenes by way of internal reactions of benzylated and benzoylated carbohydrate derivatives.

"Internal" C-glycosylarenes [e.g., (2R,3S,3aS,9bR)-3,3a,5,9b-tetrahydro-3-methoxy- and -3,7-dimethoxy-2-methoxymethyl-2H-furo[3,2-c][2]benzopyran] were prepared by intramolecular reactions of 2-O-benzyl derivatives of methyl 3,5-di-O-methyl-D-xylofuranoside (2) and their conversion into authentic C-glycosylated aromatic systems was investigated. The auxiliary benzylic linkage could not be cleaved by hydrogenolysis; isochroman derivatives (e.g., (3S)-3,4-dihydro-3-[(1R,2R)-2-hydroxy-1,3-dimethoxypropyl]-5-metho xy-1H- 2-benzopyran) were obtained under these conditions. However, oxidation of the primary benzylic position with ruthenium tetraoxide gave the corresponding lactone (a dihydroisocoumarin derivative, e.g., (2R,3S,3aS,9bR)-2,3,3a,9b-tetrahydro-3,7-dimethoxy-2-methoxy - methyl-5H-furo[3,2-c][2]benzopyran-5-one) which could be opened by saponification, thereby leading to a stereochemically unique C-glycosylbenzoic acid derivative. The same type of lactone was obtained directly from a derivative of 2 bearing a sufficiently reactive benzoyl group at O-2 (3,5-dimethoxybenzoyl); this process provides a useful approach to a heterocyclic system present in a variety of natural products. In related studies, the 2-O-phenyl substituent was found to be much less reactive than the 2-O-benzyl group in intramolecular Friedel-Crafts reactions of 2-O-substituted glycofuranosides. The first examples of successful internal C-arylation in the pyranoid series were achieved from 2-O-(3-methoxybenzyl)-D- mannopyranosides; the resulting "internal C-glycosides" [( 2R,3S,4S,4aS,10bS)-2,3,4,4a,6,10b- hexahydro-3,4,8-trimethoxy-2-methoxymethylpyrano[3,2-c][2]benzopyr an and 3,4-bis(benzyloxy)-2-benzyloxymethyl-8-methoxy analog] contain a heterocyclic skeleton closely related to that of the natural product bergenin.

Alkylation↗
PubMed1984

Effects of angiographic contrast medium on left ventricular function: evaluation by contrast angiography and radionuclide angiography.

The purpose of this study was to evaluate the effect of angiographic contrast medium on left ventricular (LV) function in 26 patients undergoing diagnostic cardiac catheterization. Beat-by-beat analysis during contrast ventriculography showed that the ejection fraction (EF) was lower in the last beats than in the first beats (P less than .02). Radionuclide angiograms were obtained the day before, as well as 15 to 65 minutes after catheterization, which included contrast ventriculography and coronary anteriography. The EF by radionuclide angiography was lower after catheterization than before (43 +/- 14% vs 47 +/- 17%, P less than .01). The EF decreased by greater than or equal to 5% in 11 of the 26 patients (42%) after catheterization. The decrease in EF in some patients was observed up to 65 minutes after catheterization and was not associated with symptoms or ST-T changes. The EF decreased in only one of nine patients who received nitroglycerin during catheterization, whereas it decreased in 10 of 17 patients who did not receive nitroglycerin (P less than .05). The EF decreased in 9 of 14 patients (64%) who had normal resting LV function, whereas it decreased in only 2 of 12 patients (17%) who had abnormal resting function (P less than .05). Thus, contrast material may depress LV function up to 1 hr and is more frequent in patients with normal resting EF. The use of nitroglycerin during catheterization may mask this effect.

Adult↗
PubMed1984

Effect of intra-aortic balloon pumping on left ventricular function: evaluation by radionuclide ventriculography.

The purpose of this study was to evaluate the effect of intra-aortic balloon pumping (IABP) on left ventricular (LV) performance in ten patients (group I) with pump failure and in ten patients (group II) with angina pectoris. Left ventricular ejection fraction, regional ejection fraction, systolic chordal shortening and volumes were measured by first-pass radionuclide ventriculography using a computerized multicrystal gamma camera. Patients in group I had significantly lower ejection fractions, regional ejection fractions and systolic chordal shortening and larger end-diastolic and end-systolic volumes than patients in group II, both on (p greater than 0.0001) and off (p less than 0.004) IABP. With IABP, the ejection fraction increased by 0 +/- 0.5% in group I and 5.8 +/- 1.7% in group II (p = 0.004); the regional ejection fraction increased by 0.1 +/- 0.9% in group I and 10.3 +/- 2.1% in group II (p = 0.0004); the systolic chordal shortening increased by 1.1 +/- 0.6% in group I and 6.5 +/- 1.1% in group II (p = 0.0006); the end-diastolic volume decreased by -0.4 +/- 6% in group I and -14 +/- 4% in group II (p = 0.07); and the end-systolic volume decreased by -1 +/- 6% in group I and -21 +/- 5% in group II (p = 0.02). Thus, IABP results in improvement in left ventricular performance in patients with angina pectoris but not in patients with pump failure.

Aged↗
PubMed1984

Correlation between changes in R wave amplitude and left ventricular volume induced by rapid atrial pacing.

To examine the Brody effect in humans, we studied 15 patients by means of coronary sinus pacing. We measured left ventricular (LV) volumes from the cardiac output (measured by the thermodilution technique) and LV ejection fraction (measured by radionuclide ventriculography). Pulmonary blood volume was determined by means of cardiac output and mean pulmonary transit time. In six patients, pacing was performed at two different rates, resulting in 21 pacing measurements. The heart rate increased with pacing from 73 +/- 11 to 119 +/- 19 bpm (mean +/- standard deviation, p less than 0.001). The end-diastolic volume (EDV) and the end-systolic volume (ESV) decreased with pacing (p less than 0.001 each). The R wave amplitude decreased with pacing (1.44 +/- 0.63 mV control vs 1.32 +/- 0.58 mV with pacing; p less than 0.01). R wave amplitude decreased in 19 of the 21 pacing studies (90%); EDV and ESV decreased in all 21 pacing studies, and pulmonary blood volume decreased in 14 of the 15 pacing studies (93%) performed in 11 patients. There was a significant correlation between the percentage of change in R wave amplitude with the percentage of change in EDV (r = 0.54, p less than 0.01) and with the percentage of change in ESV (r = 0.54, p less than 0.01). These results, therefore, validate Brody's hypothesis and indicate that changes in LV volumes affect the R wave amplitude.

Adult↗
PubMed1984

Thallium-201 myocardial scintigraphy and left ventricular function at rest in patients with rest angina pectoris.

The purpose of this study was to examine the rest thallium-201 perfusion pattern during angina-free periods in 40 patients with rest angina pectoris secondary to coronary artery disease (greater than or equal to 70% diameter narrowing). Seventeen patients had previous Q wave myocardial infarction. The perfusion defects were considered fixed or reversible, depending on the absence or presence of redistribution in the 4-hour delayed images. There were 40 perfusion defects (26 fixed and 14 reversible) in 27 patients whereas 13 patients had normal scans. Reversible perfusion defects were present in 10 patients (25%). Of the 26 fixed perfusion defects, 17 did not have corresponding Q waves. Occluded vessels (63%) had more perfusion defects than vessels with subtotal occlusion (30%) (p less than 0.01). The perfusion defect size was larger in patients with lower ejection fraction than in patients with higher ejection fraction. We conclude: (1) perfusion defects are common in patients with rest angina and are reversible in 25% of patients indicating reduced regional coronary blood flow; (2) the degree of stenosis affects the presence of perfusion defect; (3) fixed defects may be present without corresponding Q waves; and (4) global left ventricular function is related to the size of perfusion defects.

Adult↗