Artificial antigens. Synthetic carbohydrate haptens immobilized on crystalline bacterial surface layer glycoproteins. The crystalline surface-layer glycoproteins of Clostridium thermohydrosulfuricum L111-69, Bacillus stearothermophilus NRS 2004/3a and Bacillus alvei CCM 2051 were used for immobilization of spacer-linked blood group A-trisaccharide (alpha GalNAc(1----3)[alpha Fuc(1----2)]beta Gal) and of the spacer-linked, tumor-associated T-disaccharide [beta Gal(1----3)alpha GalNAc]. The immobilization involved the glycan portions of surface-layer glycoproteins. Different activation methods were used, namely, periodate oxidation, or treatment with epichlorohydrin or divinyl sulfone, followed by coupling of the hapten under appropriate conditions. The resulting conjugates are useful for assessing the application potential of haptenated surface layer preparations as carrier/adjuvants for the induction of immunity to poorly immunogenic molecules.


Structural elucidation of the capsular polysaccharide of E. coli serotype K47. The capsular polysaccharide from Escherichia coli K47 was investigated using mainly methylation analysis and 1H and 13C NMR spectroscopy and shown to have the following repeating unit: [Formula: see text]


Structural variability in the glucuronoxylomannan of Cryptococcus neoformans serotype A isolates determined by 13C NMR spectroscopy. Cryptococcus neoformans, the etiologic agent of cryptococcal meningoencephalitis, produces glucuronoxylomannan (GXM) as the major capsule component. Purified GXMs obtained from eight serotype A isolates of C. neoformans were treated by ultrasonic irradiation and then O-deacetylated prior to their comprehensive chemical analysis by GLC, GLC-MS, and 13C NMR spectroscopy. The average xylose: mannose: glucuronic acid molar ratio of the eight isolates is 1.96 +/- 0.25: 3.00: 0.58 +/- 0.10. Methylation analyses and 13C NMR spectroscopy show a general structure for GXM that is comprised of a linear (1----3)-alpha-D-mannopyranan substituted with beta-D-GlcpA and with beta-D-Xylp at O-2. Variable quantities of unsubstituted (1----3)-alpha-D-Manp were observed between the eight isolates studied. In several isolates some of the (1----3)-alpha-D-Manp residues are disubstituted with beta-D-GlcpA at O-2 and with beta-D-Xylp at O-4; this type of substitution was not previously thought to occur in serotype A isolates. Heterogeneity, between isolates, in the disposition of the substituents along the mannopyranan backbone was revealed by 13C NMR spectroscopy. The eight isolates, and three isolates previously studied, were each assigned to one of four distinct groups based on the 13C NMR chemical shifts of the anomeric carbons. Six of the eleven isolates gave identical spectra (Group I). The six major anomeric resonances from Group I were assigned to specific glycosidic linkages present in GXM. The remaining five isolates gave more complex spectra that are indicative of additional linkages and comprise the remaining three groups. Three of these five isolates contain substantial amounts of linkages previously thought to be distinctive of serotypes B and C, i.e., Manp residues that are 4-O-glycosylated with beta-D-Xylp. Methylation analyses only predicted an average repeating unit, whereas 13C NMR spectroscopy demonstrated that GXM from each isolate may be categorized into four groups by the occurrence of distinct sequences of carbohydrate residues.


Synthesis of disialoganglioside GD1 alpha and its positional isomer. The first total syntheses of disialoganglioside GD1 alpha and its alpha 2-6 positional isomer are described. Suitably protected pentasaccharide derivatives, derived from known pentasaccharide precursors, were selected as the glycosyl acceptors. Using our facile method of stereoselective alpha-glycosidation of sialic acid, these acceptors were coupled with methyl (methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-2-thio-D-glycero-D-galact o-2- nonulopyranosid)onate in acetonitrile medium in the presence of dimethyl(methylthio)sulfonium triflate (DMTST) to get the desired di-alpha-sialyl hexasaccharide derivatives in moderate yields. The hexasaccharide trichloroacetimidates upon coupling with (2S,3R,4E)-2-azido-3-O-benzoyl-4-octadecene-1,3-diol gave the corresponding beta-linked sphingosine glycosides in high yields. These were subjected in sequence to selective reduction of the azido group and coupling of the thus formed amino group with octadecanoic acid, O-deacylation, and saponification of the methyl ester groups to obtain ganglioside GD1 alpha and its positional isomer having the main-chain residue 2-6 linked.


Effects of drugs on body venous tone, as reflected by mean circulatory filling pressure. The venous system is supremely important in the control of cardiac output. Drugs which affect the venous system have profound effects on haemodynamics. This review comments on the methods available for the determination of venous compliance, resistance, and unstressed volume and describes the mean circulatory filling pressure (MCFP) technique, its usefulness and limitations. The MCFP technique involves the measurement of central venous pressure during brief (5-7 s) circulatory arrest. Mathematically, MCFP is inversely proportional to vascular compliance while experimentally, it is a primary determinant of venous return. The MCFP technique provides a reproducible and relatively non-traumatic means for the estimation of body venous tone in conscious and anaesthetised animals. Drugs examined by this technique include alpha and beta adrenoceptor agonists and antagonists, ganglionic blockers, vasoactive peptides (endothelin, vasopressin, angiotensin, neuropeptide Y), and vasodilators (hydralazine, nitroprusside, glyceryl trinitrate, calcium antagonists, and MCI-154).


Role of adenosine receptor activation in myocardial infarct size limitation by ischaemic preconditioning. The aims were to examine the role of adenosine receptors in the mechanism of preconditioning in a chronic rabbit model of myocardial infarction; to assess whether the preconditioning effect is blocked by an adenosine receptor antagonist, 8-phenyltheophylline; and to determine whether an adenosine A1 receptor agonist, R(-)N6-2-phenylisopropyl adenosine (R-PIA), mimics infarct size limitation by preconditioning. Myocardial infarction was induced in male rabbits by occlusion of the left coronary artery for 30 min, which was followed by 72 h reperfusion. Before the 30 min ischaemia, rabbits were subjected to one of the following six protocols: (1) untreated control; (2) intravenous injection of 8-phenyltheophylline; (3) preconditioning with 5 min ischaemia; (4) pretreatment with 8-phenyltheophylline plus preconditioning; (5) intravenous injection of R-PIA; or (6) R-PIA plus atrial pacing (240.min-1). Infarct size and area at risk were determined by histology and fluorescent particles, respectively. Preconditioning significantly limited infarct size, normalised as a percent of area at risk (%IS/AR), to 19.2 (SEM 2.3)% v control value of 46.5(2.8)%. 8-Phenyltheophylline alone did not modify the %IS/AR, but its injection before preconditioning attenuated the preconditioning effect such that IS/AR = 34.4(2.5)%. While R-PIA did not achieve statistically significant myocardial salvage, R-PIA plus atrial pacing limited infarct size to 33.7(3.0)% (p<0.05 v control). The R-PIA group had severe hypotension and their infarct sizes were inversely correlated with diastolic blood pressure at reperfusion. There was no such correlation in the R-PIA plus pacing group in which bradycardia and hypotension induced by R-PIA were attenuated by atrial pacing. The infarct size limiting effect of preconditioning was attenuated by 8-phenyltheophylline, and pretreatment with R-PIA was able to limit myocardial infarct size when severe hypotension was avoided by atrial pacing. These findings suggest that adenosine receptor activation plays a crucial role in the mechanism of preconditioning.


Pretreatment with buflomedil enhances ventricular function by reducing the dysfunctional area after transient coronary artery occlusion. The aim was to evaluate whether buflomedil (a drug used to treat peripheral vascular disease and which has a number of pharmacological actions potentially beneficial to dysfunctional myocardium) would preserve myocardial function after transient coronary artery occlusion followed by reperfusion. The physiological response to a 15 min balloon occlusion of the left anterior descending coronary artery followed by 1 h of reperfusion was monitored in 17 placebo treated dogs and compared with that of 15 dogs which received 10 mg.kg-1 of buflomedil. Buflomedil or its vehicle were given intravenously. Myocardial blood flow was assessed with radiolabelled microspheres and cardiac function was evaluated with quantitative contrast left ventriculography. Buflomedil did not affect baseline haemodynamic variables or contractile function. At the end of occlusion, there was no difference between dogs receiving vehicle compared with those receiving drug with respect to ejection fraction [33(SD 11)% v 34(11)%] or transmural blood flow [0.23(0.11) v 0.28(0.14) ml.g-1 x min-1]. However, at 30 min after reperfusion, ejection fraction was 89% of normal in the buflomedil group compared with 69% of normal in the placebo group (p < 0.03). This difference was sustained 60 min after reperfusion, and was due in part to slightly enhanced flow during reperfusion and a decrease in the dysfunctional area (16 compared with 28 chords lower than -2 SD from the mean, p < 0.04) in the hearts of dogs receiving buflomedil. Areas at risk were equivalent (15.9% and 15.8% of the left ventricle, respectively). The results suggest that buflomedil and agents with similar modes of action may be beneficial in preserving ventricular function after transient ischaemia followed by reperfusion.


Regional interaction and its effect on patterns of myocardial segmental shortening and lengthening during different models of asynchronous contraction in the dog. The aim was to examine the effect of asynchrony and regional myocardial interaction on the pattern of segmental contraction and relaxation. Three models of asynchrony were produced. Firstly the left anterior descending artery was abruptly occluded for 60 s. Secondly, the same artery was gradually occluded to produce four degrees of ischaemia based on the severity of the mechanical dysfunction. Finally, asynchrony was created by infusing isoprenaline (0.04 microgram.ml-1) into the left circumflex artery. Twelve anaesthetised beagles, weighing 16-21 kg, were used for the study. The patterns of contraction and relaxation were characterised by analysing the phases of shortening and lengthening, the peak lengthening rate (dL/dt), and the timing from the onset of systole to minimum systolic length. A consistent pattern of shortening and lengthening was evident during all three models of asynchrony. There were reciprocal relations between the extent of isovolumetric shortening in the normal segment and in the abnormal segment, and on occasion between the extent of isovolumetric shortening in the normal segment and the extent of isovolumetric lengthening in the same segment. Normal segments that showed minimal shortening or even some lengthening during isovolumetric systole tended to shorten beyond ejection, while segments that shortened significantly during isovolumetric contraction, lengthened earlier. Despite no change in isovolumetric shortening, segments also shortened after ejection when the opposite segment lengthened in late systole and early diastole. The pattern of shortening and lengthening depends on the path of contraction or on its entire loading pattern throughout systole. It is also possible that during early isovolumetric systole a segment can either be unloaded or preloaded by an opposing segment.


Antiarrhythmic effects of preconditioning in anaesthetised dogs and rats. The aim was to determine the relationship of the duration of short coronary artery occlusions and of the reperfusion period to the extent of the antiarrhythmic effect of preconditioning. A prolonged occlusion of a coronary artery in 102 anaesthetised rats and 55 anaesthetised dogs was preceded by a variable number of preconditioning coronary artery occlusions, of varying duration and with variable reperfusion periods between them and the prolonged occlusion. Preconditioning in both species reduced the severity of ischaemia induced arrhythmias, epicardial ST segment changes, and alterations in the degree of inhomogeneity of conduction during a subsequent prolonged coronary artery occlusion, provided that the reperfusion time was less than 30 min (in rats) and 1 h (in dogs). This antiarrhythmic effect of preconditioning was marked; eg, in dogs following two preconditioning occlusions survival from a combined ischaemia-reperfusion insult was 40% (cf, 0% in the controls). Short preconditioning periods of myocardial ischaemia protect the myocardium against the arrhythmogenic effects of a more prolonged occlusion. The optimum time for this preconditioning occlusion in rats is 3 min and protection is still apparent 30 min later. In dogs, the protective effect is especially clear with two short (5 min) coronary artery occlusions. The protection in this species lasts for less than 1 h.


Effect of adrenaline on cardiac force-interval relationship. The aim was to test the hypothesis that adrenaline affects the force-interval processes. The force-interval processes were studied in eight guinea pig papillary muscles (isometric force) and five anaesthetised dogs with atrioventricular block (maximum rate of rise of left ventricular pressure, LVdP/dtmax). The contractility indices were measured during pacing sequences in which a steady state was interrupted after a variable interval by a premature beat followed by an immediate return to the steady state. The relationship between contractility of the premature beat and the preceding interstimulus interval displays an approximately monoexponential initial rising phase, ie, mechanical restitution. With increasing adrenaline dosage in the isolated preparation there was always a significant increase in the force, and in its rate of rise with interval in some cases. Adrenaline had a variable accelerating effect on the time course of this mechanical restitution in isolated papillary muscles, but no effect in the dog preparation. In the isolated preparation adrenaline also slowed the decay in potentiation of the two beats immediately following the premature contraction. A slope of the relationship between the contractility of the second potentiated beat and that of the first was thus increased. This difference was not apparent in the intact preparation. The speeding up of mechanical restitution by adrenaline may be interpreted as reflecting the time course of reavailability of contractile activator. The slope of the relationship of contractility to that of the previous beat during the decay of postextrasystolic potentiation may be interpreted as the recirculation fraction of contractile activator; this is increased by adrenaline. However, in addition, adrenaline exerts an inotropic effect by another mechanism.


Evaluation of reperfusion strategy for the globally ischaemic rat heart. Recovery of function and energy metabolism. The sequelae of myocardial ischaemia can in principle be alleviated by repeated reperfusion, but the accumulation of adenosine monophosphate (AMP) and the loss of interstitial adenosine may lead to adenylate depletion. Repeated oxidative stress could predispose the heart to reperfusion injury. The aim of this study was to investigate the effect of intermittent reperfusion on myocardial energetics and postischaemic function. Isolated retrogradely perfused rat hearts were subjected to 20 min ischaemia, this being continuous in group I while the hearts in group II were reperfused for three 2 min periods at 5 min intervals. Function and energy metabolism were evaluated during the postischaemic reperfusion. Considerable efflux of adenosine compounds was seen during the final reperfusion, this being greater in group I than in group II, at 6.6(SEM 0.9) v 2.0(0.4) mumol.g-1 dry weight (p < 0.01). Tissue AMP, inorganic phosphate, and adenosine catabolites were higher in group I than in II after the ischaemic insult (p < 0.02), and ATP was higher in group II at the end of the final reperfusion (p < 0.05). All the hearts recovered; however, in group I the rate-pressure product was lower than in group II. Repetitive reperfusion, although short in duration, is beneficial in ischaemia in terms of lower adenylate loss and better postischaemic recovery. This should be taken into consideration when designing clinical reperfusion interventions.


Role of nitric oxide synthesis in the regulation of coronary vascular tone in the isolated perfused rabbit heart. The aim was to study the effects on coronary vascular tone of three inhibitors of nitric oxide (NO) synthesis. Studies were performed on isolated perfused hearts of 74 male New Zealand White rabbits fed normal laboratory diet. Resting coronary perfusion pressure was increased to 40-60 mm Hg with the thromboxane mimetic 9,11-dideoxy-9 alpha,11 alpha-methanoepoxy prostaglandin F2 alpha (U46619). The effects of NG-monomethyl-L-arginine (L-NMMA), N-iminoethyl-L-ornithine (L-NIO), and NG-nitro-L-arginine methyl ester (L-NAME) (0.3-300 microM) on resting coronary perfusion pressure were determined. The effects of these compounds, at concentrations that increased the resting perfusion pressure to a similar extent, on the fall in perfusion pressure induced by acetylcholine (0.1 microM) and glyceryl trinitrate (1 microM) were also investigated. In these studies the resting perfusion pressure was maintained at 40-60 mm Hg by reducing the concentration of U46619. L-NMMA, L-NIO, and L-NAME induced concentration dependent increases in resting coronary perfusion pressure (n = 3-9, p < 0.05). L-NAME had the greatest potency and efficacy, increasing the resting pressure by 48.0(SEM 9.6) mm Hg at 30 microM. L-NIO and L-NMMA increased perfusion pressure by 27.3(3.0) and 19.5(5.8) mm Hg respectively at the maximum concentration studied (300 microM). However, at concentrations that were equieffective on resting perfusion pressure (15 mm Hg increase), L-NMMA (100 microM), but not L-NIO (25 microM) or L-NAME (4 microM), significantly inhibited the fall in pressure induced by acetylcholine by 57.2(5.0)%, n = 6, p < 0.05. This effect of L-NMMA++ was attributed to a shorter duration of fall and was reversed by L-arginine (300 microM). L-NMMA (100 microM) and L-NIO (25 microM) potentiated the effect of glyceryl trinitrate by increasing the peak fall in perfusion pressure by 75.6(11.0)% and 68.8(24.1)% respectively (n = 6 for each, p < 0.05). The differential effects of the three inhibitors on resting coronary perfusion pressure and the acetylcholine induced fall in coronary perfusion pressure suggest that basal and stimulated NO synthesis may be differentially regulated. Reduction in the synthesis of endogenous NO by these compounds potentiates the glyceryl trinitrate induced fall in perfusion pressure, which may have important clinical implications.


Activation of bovine endothelial thromboxane receptors triggers release of prostacyclin but not EDRF. The aim was to examine the capacity of U46619 (a stable thromboxane A2 mimetic) to mediate release of endothelium derived relaxing factor (EDRF) from bovine aortic endothelial cells, and compare the response to the U46619 dependent release of prostacyclin (PGI2). Bovine aortic endothelial cells (AG4762) were cultured in vitro on microcarrier beads, which were then loaded onto a column and perfused. The cells were challenged with U46619, bradykinin, or the Ca2+ ionophore ionomycin in the perfusate, and measurements made of the release of 6-oxo-PGF1 alpha (the stable hydrolysis product of PGI2, measured by radioimmunoassay) and EDRF (bioassay). Cells were also cultured on glass cover slips, loaded with Fura 2-AM, and measurements made of the rise in intracellular Ca2+ after challenge with U46619, bradykinin or ionomycin. U46619 triggered release of 6-oxo-PGF1 alpha but not EDRF from AG4762 cells, contrasting with bradykinin which released both 6-oxo-PGF1 alpha and EDRF. Ionomycin had little or no capacity to mimic and trigger release of 6-oxo-PGF1 alpha, although ionomycin mediated large increases in intracellular Ca2+. In contrast, staurosporine (a putative inhibitor of protein kinase C) substantially inhibited the U46619 and bradykinin dependent release of 6-oxo-PGF1 alpha. In contrast to bradykinin linked receptors on AG4762 endothelial cells, which are coupled to the release of both prostacyclin and EDRF, activation of thromboxane A2 receptors on these cells selectively triggers release of PGI2 but not EDRF. Further, based on the distinct effects of ionomycin and staurosporine, it appears that agonist stimulated PGI2 release from these cells is mediated predominantly by protein kinase C, rather than by rises in intracellular Ca2+. This observation contrasts with previously described mechanisms of PGI2 release from endothelium obtained from other sources.


Hypoxic preconditioning of ischaemic canine myocardium. The aim was to test whether a brief period of non-ischaemic hypoxia can precondition myocardium. 60 anaesthetised adult mongrel dogs of either sex underwent 60 min occlusion of the left anterior descending coronary artery, followed by 5 h reperfusion. In treated groups, hearts were either preconditioned with 5 min coronary perfusion with hypoxic blood [O2 content 9.2(SEM 0.6) ml.litre-1] or 5 min occlusion followed by a 10 min reperfusion period prior to 60 min occlusion. The effect of these treatments on myocardial infarct size and regional contractile function was assessed. Infarct size, determined by tetrazolium staining, as a percentage of anatomical area at risk was markedly decreased in hypoxia preconditioned hearts, at 7.2(1.8)% v 22.4(4.6)% in controls (p < 0.01), but did not differ from ischaemia preconditioned hearts [4.6(1.7)%; p < 0.01 v control]. Anatomical area at risk, expressed as a percentage of left ventricular mass, and collateral blood flow to the inner two thirds of the ischaemic wall did not differ among the groups. Regional contractile function was depressed following ischaemic preconditioning but not following hypoxic preconditioning. During reperfusion following 60 min occlusion, marked paradoxical systolic lengthening was evident in ischaemia preconditioned and control hearts but not in hypoxia preconditioned myocardium. Five minutes of hypoxic and ischaemic preconditioning were equipotent in preventing infarction, whereas ischaemic preconditioning caused a greater decrement in postischaemic contractile function.


Cardiac A2 adenosine receptors--influence of ischaemia. The aim was to detect cardiac A2 adenosine receptors through radioligand binding, and to assess the effect of ischaemia on these receptors. Isolated working rat hearts were subjected either to aerobic perfusion or to global ischaemia. A membrane fraction was prepared from ventricular tissue, and 3H-5'-N-ethylcarboxamide adenosine (NECA) binding was determined in the presence of N6-cyclopentyl adenosine (CPA). A2 binding was calculated as the fraction of NECA binding displaced by 100 microM CPA but not displaced by 50 nM CPA. Analysis of A2 NECA binding according to single binding site model yielded Kd = 22.0 nM, Bmax = 34.0 fmol.mg-1 in control hearts; Kd = 49.7 nM, Bmax = 44.3 fmol.mg-1 in hearts subjected to 30 min ischaemia (p < 0.05 for difference in Kd). In the control group a two site model provided a significantly (p < 0.05) better fit (Kd = 5.6 and 183.7 nM, Bmax = 9.5 and 64.4 fmol.mg-1 for the high and low affinity sites respectively). The high affinity component of A2 NECA binding disappeared in the presence of the GTP analogue guanyl-5'-yl imidodiphosphate, suggesting the existence of multiple coupling states of the receptor. In the ischaemic group no significant improvement in data fitting was obtained with the two site model. The results provide evidence of the existence of cardiac A2 adenosine receptors. Ischaemia modifies receptor properties and appears to affect chiefly the high affinity component of A2 binding, possibly by preventing receptor interaction with membrane G proteins.


Coronary aneurysms: a case report and review. This case report describes a large coronary aneurysm and poses questions regarding management. The discussion that follows addresses what is known about the natural history of and options for management of coronary aneurysms.


Left main coronary artery aneurysm following percutaneous transluminal angioplasty: a report of a case and review of the literature. Since its introduction in 1977, the number of PTCAs and its indications have grown. Along with more frequent usage, newer complications have been reported. Aneurysm of left main coronary artery is rare. This report describes the formation of a new non-obstructing aneurysm in the left main coronary artery after PTCA of left circumflex artery. The patient has had 7 yr of follow-up with a benign clinical course.


Coronary artery aneurysm formation following directional coronary atherectomy. Directional coronary atherectomy has recently become available to treat coronary stenosis by excision and removal of tissue. The optimal depth of resection by this method has not been determined and complications have occurred. This report describes the formation of a coronary aneurysm at an atherectomy site in an asymptomatic patient, a finding not reported previously.


Case report: formation of vessel aneurysm after stand alone coronary excimer laser angioplasty. Formation of aneurysms in coronary arteries can be observed following percutaneous transluminal balloon angioplasty but has not been reported previously after coronary excimer laser angioplasty in humans. Stand alone coronary excimer laser angioplasty was performed in a 49-year-old man with a 75% left anterior descending artery stenotic lesion and exertional angina, documenting a good angiographic result postintervention. Control angiography 6 months after the procedure revealed an aneurysm distal to a 90% restenosis in the area of ablation.


Aneurysmal formation after successful pulsed laser coronary angioplasty. We report two cases of coronary artery aneurysmal formation as long term consequences of Ho:YAG and excimer laser coronary angioplasty. This case report suggests that pulsed laser angioplasty may result in severe vessel wall damage that may lead ultimately to aneurysmal formation.


An unusual cause of recurrent angina two years after coronary artery bypass grafting: fistula between internal mammary artery graft to pulmonary vasculature. A 55-year-old man developed recurrent angina pectoris 2 years after coronary artery bypass grafting. Cardiac catheterization demonstrated that the venous grafts were patent, but selective left internal mammary angiogram showed multiple fistulous connections between the internal mammary artery and the pulmonary vasculature of the left upper lobe. After surgical correction of the fistula, the angina resolved. Only three previous cases of acquired internal mammary artery graft fistulas draining to the pulmonary vasculature have been described. The etiology, clinical presentation, and management of an internal mammary artery fistula to the pulmonary vasculature are discussed.


Angioplasty of tortuous left internal mammary artery: description of a new technique. Angioplasty of the distal anastomosis site of a markedly tortuous left internal mammary artery (LIMA)--anterior descending artery is described. Tortuosity of LIMA was negotiated using the Tracker-Seeker catheter guidewire system. Angioplasty was then performed using the conventional technique.


Detachable balloon embolization of an iatrogenic aortocoronary arteriovenous fistula combined with aortocoronary bypass PTCA: a case report. Coronary artery bypass surgery was performed in a 54-year-old male, and one of the grafts was inadvertently sutured to the vena cordis magna instead of to the left anterior descending coronary artery (LAD). Four years later the patient observed a progression of symptoms including episodes of angina pectoris at rest. Coronary angiography showed stenosis of one of the bypass grafts and notable dilatation of an iatrogenic arteriovenous (A-V) fistula. The stenosed bypass graft was dilated and the A-V fistula occluded by use of a detachable balloon. Embolization was performed rather than surgery, as the LAD was found to be a poor surgical target. The patient's symptoms improved after the procedure; he was followed for 18 months during which time his condition remained stable.


Combined PTCA and microcoil embolization of a left internal mammary artery graft. Internal mammary arteries are increasingly common conduits for coronary revascularization. Although infrequent, cardiologists are faced with a number of technical failures. We describe a case of combined PTCA to a LIMA insertion stenosis, along with coil embolization of a large unligated intercostal side branch. A complication of embolization is described along with angiographic follow-up.


Modified "kissing" atherectomy procedure with dependable protection of side branches by two-wire technique. We report a new two-wire atherectomy technique for side branch protection. Newer, more resilient wire designs are capable of withstanding cutting forces of the Simpson atherocath device. This technique expands atherectomy application to lesions previously excluded from atherectomy as high risk lesions.


Clinical results and quality of life after percutaneous transluminal coronary angioplasty: a preliminary report. To evaluate the effect of percutaneous transluminal coronary angioplasty (PTCA) on quality of life, data on symptomatic status, functional capacity, life satisfaction, and psychological wellness were collected on 102 patients at 1 day pre-PTCA and 2 months post-PTCA, and on the first 50 of these patients at 10 months post-PTCA. There were highly significant changes (p < 0.001) in all quality of life measures between pre-PTCA and the 1st follow-up measurements. No further significant changes occurred in these measures between the 1st and 2nd follow-up measurements, indicating that the initial improvement in quality of life was sustained over this period. Data on primary success rate, complications, and pre- and post-PTCA risk factor scores are also reported.


Does the AHA/ACC task force grading system predict outcome in multivessel coronary angioplasty? To assess the ACC/AHA task force grading system as a predictor of outcome in patients undergoing multivessel percutaneous transluminal coronary angioplasty we analyzed all failures (residual stenosis > 50%, Q-wave myocardial infarction, coronary artery bypass grafting during hospitalization, or death) in 97 patients with 328 stenoses. There were 70 males and 27 females; 60 patients had stable angina, and 37 had unstable angina. The mean number of lesions dilated per patient was 3.4 (range 2-8). The mean preangioplasty percent luminal diameter narrowing was 80 +/- 14%. Thirty-eight stenoses were AHA/ACC classification type A, 192 type B, and 98 type C. One hundred twenty-eight lesions were located in the left anterior descending artery or its distribution, 89 in the left circumflex, 96 in the right coronary artery, and 15 in other vessels. Procedural success (< 50% residual diameter narrowing and no major ischemic complications) was achieved in 266 lesions (81.1%). Major ischemic complications (death, myocardial infarction, or emergency bypass surgery) occurred in 8 patients (8.2%) and in-hospital mortality was 2%. Analysis on a per stenosis basis demonstrated 84% success in type A, 89% in type B, and 64% in type C (p < 0.0001). When type B was divided into type B1 (1 type B characteristic) and type B2 (two or more type B characteristics) the success rate was 90% vs. 88% and the complication rate was 1% vs. 2%, respectively (p = n.s.). Logistic regression analysis showed that the best single predictor of failed angioplasty was total occlusion > 3 months, followed by total occlusion < 3 months and severely angulated (> 90 degrees) segment.(ABSTRACT TRUNCATED AT 250 WORDS)


The attachments of chromatin loops to the nucleoskeleton. It is widely assumed by cell biologists that chromatin is looped by attachment to some nuclear skeleton. 'Structural' attachments might be mediated through specific sequences; these would be attached in most cells in an organism, underlying the basic structure of the mitotic chromosome and persisting throughout interphase. 'Functional' attachments might also exist, perhaps if active polymerases are attached to the skeleton and replication and transcription occur as DNA is reeled through them. Cells of different tissues--and even cells of the same tissue--would have different attachments of this type. Problems associated with demonstrating these two kinds of attachment are discussed. We find little good evidence for 'structural' attachments and explore the idea that 'functional' attachments are the only kind that exist: 'functional' attachments involving active transcription units might be stable enough to organize chromatin during both interphase and mitosis, but 'dynamic' enough to allow duplication of attached sequences without disrupting loops.


The transcribed template and the transcription loop in Balbiani rings. The active Balbiani ring (BR) genes in the giant puffs in Chironomus salivary glands can be visualized in three dimensions within the cell nucleus. The chromatin template exhibits a dynamic structure: at the RNA polymerases the template is fully extended, while in between polymerases it can be packed into a nucleosome, a nucleosome filament, or even into a thick chromatin fiber, if the spacing between consecutive polymerases is large enough. Even in its extended state, the template harbours histone H1, and most likely also the core histones, indicating that the nucleosomes unfold and refold during transcription. Apart from the transcribed template, a transcription loop contains a thin, extended, apparently flexible and nucleosome-free fiber upstream of the gene, and a loosely coiled nucleosome filament downstream of the gene.


Localization of nuclear matrix core filament proteins at interphase and mitosis. The gentle removal of chromatin uncovers a nuclear matrix consisting of two parts: a nuclear lamina connected to the intermediate filaments of the cytoskeleton and an internal matrix of thick, polymorphic fibers connecting the lamina to masses in the nuclear interior. This internal nuclear matrix can be further fractionated to uncover a highly branched network of 9 nm and 13 nm core filaments retaining some enmeshed bodies. The core filament network retains most of the nuclear RNA, as well as the fA12RNP antigen, and may be the most basic or core element of internal nuclear structure. One high molecular weight protein component of the core filament network, the H1B2 antigen, is normally masked in the interphase nucleus and is uncovered as the chromatin condenses at mitosis. This protein is associated with a fibrogranular network surrounding and connected to the chromosomes. The core filament-associated fA12 antigen also becomes associated with this perichromosomal network. We propose that the core filament nuclear matrix structure may not completely disassemble at mitosis but, rather, that parts remain as a structural network connected to chromosomes and other mitotic structures. These mitotic networks may, in turn, serve as the core structures on which the nuclear matrices of daughter cells are built.


Three-dimensional immunogold labeling of nuclear matrix proteins in permeabilized cells. A preembedment labeling procedure is described for the three-dimensional (3D) labeling of nuclear matrix proteins in permeabilized cells. The procedure is based on the use of ultra-small (1 nm) gold particles as a marker system. This marker penetrates the nucleus more efficiently than the conventionally used 5-10 nm colloidal gold probes. Dehydration is performed by freeze-substitution to preserve the ultrastructure of the cell as optimally as possible. During freeze-substitution the samples are stained by uranyl ions to stain the cellular material throughout the resin section. The 3D gold-labeled and uranyl-stained specimen is embedded in Epon resin and semi-thin (0.2-0.5 microns) sections are made for stereo electron microscopy. The applicability of this method is illustrated by the localization of nuclear matrix-associated nuclear bodies in permeabilized interphase and mitotic HeLa cells.


Indonesia medicinal plants. III. On the constituents of the bark of Fagara rhetza (Rutaceae). (1): Alkaloids, phenylpropanoids, and acid amide. Two new phenylpropanoids, named O-geranylsinapyl alcohol (1) and O-geranylconiferyl alcohol (2), and a new acid amide, named hazeleamide (3), were isolated from the bark of Fagara rhetza (Rutaceae), an Indonesian medicinal plant from Flores Island, Indonesia. The chemical structures of 1, 2, and 3 have been elucidated on the basis of their chemical and physicochemical properties. Among the three new compounds, hazeleamide (3) was found to show a pungent taste and to exert a moderate antimalarial activity in an in vitro test system.


Studies on the chemical modification of monensin. IV. Synthesis, sodium ion permeability, and biological activity of 7-O-acyl- and 7-O-alkylmonensins. 7-O-Acyl-(4a-e) and 7-O-alkylmonensins (5a-d) were prepared from monensin (1). Their lipophilicity, sodium ion permeability in human erythrocytes, antibacterial activity and effect on rat tail artery were examined. There was a correlation between lipophilicity and sodium ion permeability as well as between lipophilicity and antibacterial activity. We also found that the compound having larger sodium ion permeability, showed stronger contraction of rat tail artery. 7-O-Benzylmonensin (5c) exhibited higher lipophilicity and larger sodium ion permeability than monensin (1) among the tested monensin derivatives. In addition, antibacterial activity and contractile effect on rat tail artery of 5c were comparable to those of 1.


Synthesis of 1,4-dihydropyridine-5-phosphonates and their calcium-antagonistic and antihypertensive activities: novel calcium-antagonist 2-[benzyl(phenyl)amino]ethyl 5-(5,5-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-1,4-dihydro-2,6-dim ethyl-4-(3-nitrophenyl)-3-pyridinecarboxylate hydrochloride ethanol (NZ-105) and its crystal structure. The effect of the 3-carboxylic-ester variation in 2,2-dimethyltrimethylene 3-alkoxycarbonyl-4-aryl-1,4-dihydro-2,6-dimethyl-5-pyridinephosphonat es (1) was investigated with relation to the calcium-antagonistic and antihypertensive activities: the analogs containing the alkyl groups of not more than 12 carbons and an amino functionality in the carboxylic-ester moiety were synthesized to be examined for biological activities. Among them, 2-[benzyl(phenyl)amino]-ethyl 5-(5,5-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-1,4-dihydro-2, 6-dimethyl-4-(3-nitrophenyl)-3-pyridine-carboxylate hydrochloride ethanol (NZ-105) showed particularly beneficial activities and was selected for further pharmacological studies and clinical development. Some aspects of the structure-activity relationships and solid-state structure of NZ-105 by X-ray crystallographic analysis were described.


Synthesis and biological activities of optical isomers of 2-(4-diphenylmethyl-1-piperazinyl)ethyl 5-(4,6-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-1,4-dihydro-2,6-dim ethyl-4-(3-nitrophenyl)-3-pyridinecarboxylate dihydrochloride (NIP-101). Six optical isomers of 2-(4-diphenylmethyl-1-piperazinyl)ethyl 5-(4,6-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-1,4-dihydro-2, 6-dimethyl-4-(3-nitrophenyl)-3-pyridinecarboxylate dihydrochloride (NIP-101, 1.2HCl.2H2O), a potent calcium antagonist, were successfully prepared by using optically active (2R,4R)-(-)- and (2S,4S)-(+)-2,4-pentanediols, and cis-2,4-pentanediol and optically active (S)-(+)-2-methoxy-2-phenylethanol. Their proton nuclear magnetic resonance investigations demonstrate that the 1,3,2-dioxaphosphorinane group is conformationally constrained around the C-P bond. Calcium-antagonistic and hypotensive activities of the optical isomers were examined and found to depend mainly on the absolute configuration at a stereogenic center in the 1,4-dihydropyridine ring rather than the configuration of the 1,3,2-dioxaphosphorinane moiety.


Synthesis and crystal structure of optically active 2-[benzyl(phenyl)amino]ethyl 5-(5,5-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-1,4-dihydro-2,6-dim ethyl-4-(3-nitrophenyl)-3-pyridinecarboxylate (NZ-105). (S)-2-[Benzyl(phenyl)amino]ethyl 5-(5,5-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-1,4-dihydro-2, 6-dimethyl-4-(3-nitrophenyl)-3-pyridinecarboxylate ((S)-NZ-105) and R isomer were synthesized through the fractional crystallization of (S)-2-Methoxy-2-phenylethyl 5-(5,5-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-1,4-dihydro-2, 6-dimethyl-4-(3-nitrophenyl)-3-pyridinecarboxylate. Calcium antagonism activity was found to reside in the S isomer from single crystal X-ray diffraction analysis.


Preparation and pharmacological evaluation of 4-(1,4-benzoquinon-2-yl)-4-phenylbutanamides as potential cerebral protective agents. A new series of 4-(1,4-benzoquinon-2-yl)-4-phenylbutanamides (2) were synthesized for evaluation of their pharmacological activities. All these compounds synthesized showed significant antilipidperoxidation (ALP) activities with brain homogenate in rats and some of them possessed a protective effect against hypobaric hypoxia in mice. Especially, a thiomorpholine derivative (2l, SUN-4757) showed a wide efficacy spectrum to a variety of experimental screening assays designed for cerebral protective agents, and it had a high LD50 value.


Novel disease-modifying antirheumatic drugs. I. Synthesis and antiarthritic activity of 2-(4-methylphenyl)benzothiazoles. A series of 2-(4-methylphenyl)benzothiazoles was synthesized and evaluated using an adjuvant-induced arthritic rat model. This class of desired compounds affecting the immune response was found using hemagglutination assay. 4-Acetoxy-2-(4-methylphenyl)benzothiazole (7m), KB-2683, was most potent in the adjuvant-induced arthritic rat model and selected for further evaluation. In contrast to nonsteroidal antiinflammatory drugs, compound 7m showed no antiinflammatory or analgesic activities. It did, however, show an immunomodulatory activity in enhanced delayed type hypersensitivity.


Synthesis and antiallergy activity of [1,3,4]thiadiazolo[3,2-a]-1,2,3-triazolo[4,5-d]pyrimidin-9(3H)-one derivatives. II. 6-Alkyl- and 6-cycloalkylalkyl derivatives. A series of 6-alkyl- or 6-(cycloalkylalkyl)-[1,3,4]thiadiazolo[3,2- a]-1,2,3-triazolo[4,5-d]pyrimidin-9(3H)-ones 1b--o was synthesized from the corresponding 1,3,4-thiadiazol-5-amines 3b--o and the antiallergic activities of the products were evaluated. Among the compounds 6-(2-cyclohexylethyl)- [1,3,4]thiadiazolo[3,2-a]-1,2,3-triazolo[4,5-d]pyrimidin-9(3H)-one 1h, whose X-ray crystallographic stereostructure is shown, was found to be a promising new antiallergic agent, which has low toxicity and dual activity as a leukotriene D4 receptor antagonist and as an orally active mast cell stabilizer.


Studies on antiinflammatory agents. I. Synthesis and pharmacological properties of 2'-phenoxymethanesulfonanilide derivatives. Various 2'-phenoxymethanesulfonanilide derivatives were synthesized and evaluated for antiinflammatory and analgesic activities. Some compounds bearing an electron-attracting substituent at the 4'-position strongly inhibited adjuvant-induced arthritis in rats and acetic acid-induced writhing syndrome in mice without causing gastro-intestinal irritation. Among them, 4'-cyano-(FK867) and 4'-acetyl-(FK3311) 2'-(2,4-difluorophenoxy)methanesulfonanilides were selected as the candidates for further development.


Synthesis and antibacterial activity of 1-substituted-methyl carbapenems. The synthesis of the 1 alpha- and the 1 beta-substituted (fluoro, cyano, hydroxy and acetoxy)-methyl carbapenems having a 2-(1,3,4-thiadiazol-2-yl)thiomethyl side chain are described, and their in vitro antibacterial activities are compared with the corresponding 1 beta-methyl carbapenems together with imipenem. The synthesis and antibacterial activity of the 1 beta-substituted (fluoro and cyano)-methyl carbapenems having 2-(1-alkyl-4-pyridinio)thiomethyl side chains are also described.


Syntheses and anti-inflammatory activity of novel oximes and O-acyloximes. Novel oximes were prepared from the corresponding aldehyde or ketone in the usual way, and a number of oxime esters, O-lauroyl, O-2-pyridinecarbonyl, O-nicotinoyl, and O-isonicotinoyl oximes were synthesized by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI)-4-dimethylaminopyridine (DMAP) method or a mixed anhydride method, in our search for potent anti-inflammatory compounds. The anti-inflammatory activity of these compounds was assessed by the carrageenan-induced paw edema assay in rats. The oximes (4, 5, and 13), O-lauroyloxime 1L, O-nicotinoyloximes (1N, 2N, 3N, and 4N), O-isonicotinoyloxime 1I, and O-2-pyridinecarbonyloxime 7P showed higher anti-inflammatory potency than aspirin, a prostaglandin cyclooxygenase inhibitor.


Studies on cerebral protective agents. II. Novel 4-arylpyrimidine derivatives with anti-anoxic and anti-lipid peroxidation activities. (2). In a search for new cerebral protective agents with anti-anoxic (AA) and anti-lipid peroxidation (ALP) activities, a series of 4-arylpyrimidines, bearing an amino moiety in the C-5 position of the pyrimidine nucleus, was synthesized and tested for AA and ALP activities. Among them, 6-methyl-5-(4-methylpiperazin-1-ylcarbonyl)-4-(3-nitrophenyl )-2-phenylpyrimidin e (41, FK360) was most effective on both assays and on arachidonate-induced cerebral edema in rats. Structure-activity relationships in regard to AA activity of this series of compounds are also discussed.


Synthesis and antiarrhythmic activity of 2,5-disubstituted 2,3-dihydro-1,2,5-benzothiadiazepin-4(5H)-one 1,1-dioxides. A series of 2,5-disubstituted 2,3-dihydro-1,2,5-benzothiadiazepin-4(5H)-one 1,1-dioxide derivatives were prepared and evaluated for the antiarrhythmic effect on ouabain-induced arrhythmias in guinea pigs. Most of the synthesized compounds showed the antiarrhythmic activity in this primary screening system. Some of the compounds with 2-(N,N-dimethylamino)ethyl, 2-(pyrrolidin-1-yl)ethyl and 2-oxo-2-(morpholin-4-yl)ethyl moieties on the 5-position of 1,2,5-benzothiadiazepin-4(5H)-one 1,1-dioxide exhibited a potent antiarrhythmic activity. The structure-activity relationship of these compounds was discussed.


Inhibitors of prostaglandin biosynthesis from Dalbergia odorifera. The root heartwood of Dalbergia odorifera T. Chen (Leguminosae) is a Chinese medicinal drug (Japanese name koshinko) used for a stagnant blood syndrome (stagnation of disordered blood; Japanese, oketsu). In addition to 10 known compounds, five new phenolic compounds, isomucronustyrene and hydroxyobtustyrene (cinnamylphenols), (+)-isoduartin (isoflavan), odoriflavene (isoflav-3-ene) and (-)-odoricarpan (pterocarpan) were isolated and their structures were elucidated on the basis of chemical and spectroscopic methods. Of the fifteen compounds isolated, cinnamylphenols, isoflavans, isoflavene and benzoic acid derivative significantly inhibited prostaglandin biosynthesis as well as platelet aggregation induced by arachidonic acid.


Studies on absorption, distribution, excretion and metabolism of ginseng saponins. VIII. Isotope labeling of ginsenoside Rb2. To clarify the pharmacokinetics of absorption, distribution and excretion of ginsenoside Rb2 (Rb2), one of the major saponins of the root of Panax ginseng, following oral administration to rats, a tritium (3H) labeling of Rb2 was examined. The C-12 position of Rb2 was labeled with 3H-sodium borohydride (3H-NaBH4) and 12-3H Rb2 and 12-3H-epi Rb2 was synthesized. This method of specific position labeling of Rb2 may be applicable to other ginsenosides. In the near future, the pharmacokinetics of Rb2 in rats may be clarified with 3H labeled Rb2.


Changes in enzymatic and membrane-adsorbing activities of sphingomyelinase from Bacillus cereus by modification with a polyethylene glycol derivative. Sphingomyelinase (SMPLC) from Bacillus cereus was modified with a polyethylene glycol (PEG) derivative, methoxypolyethylene glycol-succinimidyl succinate (ss-PEG). The molecular weight of the ss-PEG-modified SMPLC was calculated to be approx. 150 kDa by gel-filtration whereas that of the native enzyme, was 25 kDa. By this modification, the enzyme increased its thermostability and retained its hydrolytic activity toward 2-(N-hexadecanoylamino)-4-nitrophenylphosphocholine (HNP) and sphingomyelin (SM) in the mixed micelles with the surfactants such as Triton X-100 and sodium deoxycholate (SDC). However, the activity toward liposomal SM was significantly decreased, and all the enzyme activities toward bovine erythrocytes, including membraneous SM-hydrolyzing and hemolytic activities as well as the enzyme adsorption onto the erythrocyte membranes, were completely lost.


Effect of metal ions on transcription of the ada gene which encodes O6-methylguanine-DNA methyltransferase of Escherichia coli. The effect of metal ions on transcription of the ada gene of Escherichia coli which is promoted by Ada protein in the presence of methylated deoxyribonucleic acid (DNA) was examined in a reconstituted system. Their effect on the O6-methylguanine-DNA methyltransferase (MGTase) activity of Ada protein was also examined. Ag+, Cd2+, Cu2+ and Hg2+ severely inhibited transcription of the ada gene at a dose which inhibited neither transcription of the lacUV5 gene nor MGTase activity. Zn2+ inhibited both transcription and MGTase activity in the same dose range. Al3+ and Fe3+ inhibited transcription of both ada and lacUV5 genes without affecting the MGTase activity of Ada protein. Inhibitory mechanisms are discussed.


Core structure of glycyrrhizan GA, the main polysaccharide from the stolon of Glycyrrhiza glabra var. glandulifera; anti-complementary and alkaline phosphatase-inducing activities of the polysaccharide and its degradation products. The controlled Smith degradation and limited hydrolysis of glycyrrhizan GA, a representative polysaccharide with remarkable phagocytosis-enhancing activity isolated from the stolon of Glycyrrhiza glabra L. var. glandulifera Reg. et Herd. were carried out. Methylation analyses of the primary and the secondary Smith degradation products and of the limited hydrolysis product indicated that the core structural features of glycyrrhizan GA include a backbone chain composed of beta-1,3-linked D-galactose residues. Three-fifths of the galactose units in the backbone carry side chains composed of beta-1,3- and beta-1,6-linked D-galactosyl residues at position 6. Anti-complementary and alkaline phosphatase-inducing activities of the polysaccharide, periodate oxidation-reduction and the controlled Smith degradation products were investigated, and the controlled Smith degradation product showed significant activity.


Application of microdialysis for study of caffeine distribution into brain and cerebrospinal fluid in rats. The usefulness of microdialysis was examined for the chronological determination of caffeine concentration in the brain and cerebrospinal fluids (CSF) following intravenous administration of caffeine in rats. The recovery percent of caffeine by microdialysis, the concentration ratio of caffeine in the dialysate against that in the brain tissue or CSF was determined. The recovery percent was proved to be constant at 5 different steady-state plasma concentrations of caffeine (0.1-280 nmol/ml) and in different collecting periods of dialysate ranging from 30 s to 10 min. The mean recovery percent in the brain and CSF were 10.9 and 13.1%, respectively. Thus, microdialysis was proved useful for determination of drug concentration in the tissue and biological fluids with time resolution of more than 30 s. The microdialysis method was then applied for the chronological determination of caffeine concentration in the brain and CSF following intravenous bolus administration. The estimated caffeine concentration in the brain and CSF was the same as those obtained by direct determination in isolated brain and CSF, respectively. Transfer of caffeine from plasma to brain and CSF were further pharmacokinetically analyzed using a modified 2-compartment model. In this kinetic model, the transfer of caffeine between the CSF and brain was neglected, since the mutual transfer of caffeine was not detected in in vivo experiments. Calculated curves were well fitted on observed caffeine concentrations in the plasma, brain and CSF.


Plasma alpha 1-acid glycoprotein concentration in rats with chemical liver injury. The influence of liver injury on the plasma concentrations of alpha 1-acid glycoprotein (AGP) and albumin was examined in several different models of chemically-induced liver injury. The plasma AGP concentration in carbon tetrachloride (CCl4), allyl alcohol, bromobenzene, acetaminophen or N-nitrosodimethylamine-induced liver injury was increased to 2-3.5 times the normal level at 24 h after the intoxication. The plasma AGP concentration was unchanged in ethionine-induced liver injury and was markedly decreased in galactosamine-induced injury. The plasma albumin concentration was significantly decreased by the damage due to galactosamine, allyl alcohol or N-nitrosodimethylamine-induced liver injury, while no influence was observed by other hepatotoxin-induced liver injury. The plasma protein binding of propranolol was also determined in relation to the plasma concentrations of AGP and albumin in all the experimental models. Propranolol binding, expressed as bound to free ratio, showed a good correlation with the AGP concentration (r = 0.940; p < 0.001), but not with the albumin concentration.


Syntheses of 2-chloro-4-nitrophenyl beta-D-maltopentaosides with bulky modification and their application to the differential assay of human alpha-amylases. Four novel maltopentaosides, 2-chloro-4-nitrophenyl O-(6-O-p-toluenesulfonyl-alpha-D-glucopyranosyl)-(1-->4)-tris[O- alpha-D-glucopyranosyl-(1-->4)]-beta-D-glucopyranoside (4), 2-chloro-4-nitrophenyl O-[6-O-(tert-butyldimethyl)silyl-alpha-D- glucopyranosyl]-(1-->4)-tris[O-alpha-D-glucopyranosyl-(1-->4)]-beta-D- glucopyranoside (5), 2-chloro-4-nitrophenyl O-[6-deoxy-6-(phenyl)sulfonyl-alpha-D- glucopyranosyl]-(1-->4)-tris[O-alpha-D-glucopyranosyl-(1-->4)]-beta-D- glucopyranoside (10), and 2-chloro-4-nitrophenyl O-(6-deoxy-6-phthalimido-alpha-D-glucopyranosyl)- (1-->4)-tris[O-alpha-D-glucopyranosyl-(1-->4)]-beta-D-glucopyranoside (11) were synthesized. Substrates 4, 5, 10, and 11 were hydrolyzed by human pancreatic alpha-amylase (HPA) from 1.1 to 2.9-fold faster than by human salivary alpha-amylase (HSA). Taking advantage of the difference in the hydrolytic rate of 5 (2.9-fold faster), we developed a new method for the differential assay of these two human alpha-amylases.


Synthesis and antiplatelet activities of N-arylmethyl-3,4-dimethylpyrano[2,3-c]pyrazol-6-one derivatives. A series of new 1- and 2-arylmethyl-3,4-dimethylpyrano[2,3-c]pyrazol-6-one derivatives were synthesized and examined for their antiplatelet activities. Some of these compounds showed significant inhibitory activities. Among them, 1-phenylmethyl-3,4-dimethylpyrano[2,3-c]pyrazol-6(1H)-one (4a), 2-(2'-methoxyphenyl)methyl-3,4-dimethylpyrano[2,3-c]pyrazol-6(2H)- one (3e) and 2-(3'-methoxyphenyl)methyl-3,4-dimethylpyrano[2,3-c]pyrazol-6-(2H) - one (3f) were the most effective. These inhibitors acted in a concentration-dependent manner. The antiplatelet effect of compound 3f is due to the inhibition of thromboxane A2 formation and the blockade of thromboxane A2/prostaglandin endoperoxide receptor in washed rabbit platelets.


Synthesis of a dibenz[b,e]oxepin-bovine serum albumin conjugate for radioimmunoassay of KW-4679 ((Z)-11-[3-(dimethylamino)propylidene]-6,11- dihydrodibenz[b,e]oxepin-2-acetic acid hydrochloride). (Z)-11-[3-(Dimethylamino)propylidene]-2-(methoxycarbonyl)methyl-6, 11- dihydrodibenz[b,e]oxepin-9-acrylic acid (5) was prepared for application to the radioimmunoassay of KW-4679 (1, (Z)-11-[3-(dimethylamino)propylidene]-6,11-dihydrodibenz[b,e ] oxepin-2-acetic acid hydrochloride). The acrylic acid moiety in the 9-position of 5 was employed for coupling with an amino group of bovine serum albumin (BSA) to provide 17. Subsequently, the conjugate 17 was treated with aqueous NaOH to hydrolyze the terminal methoxycarbonyl group in the 2-position of the BSA conjugated 5. Antiserum raised against the antigenic BSA-conjugate 4 finally obtained was specific for 1.


Biologically active complexes of nickel(II), copper(II) and zinc(II) with Schiff-base ligand derived from the reaction of 2-aminopyridine and pyrrol-2-carboxaldehyde--their synthesis and characterisation. A new Schiff-base ligand N-(2'-pyrrylmethylidene)2-aminopyrimidine derived from the reaction of 2-amino pyrimidine and pyrrol-2-carboxaldehyde and its nickel(II), copper(II) and zinc(II) complexes have been synthesised and characterised on the basis of elemental analysis, molar conductance, infrared, electronic and proton nuclear magnetic resonance (1H-NMR) and magnetic susceptibility data. The ligand and its complexes when screened for antibacterial activity against bacterial species such as, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae. In all cases, the activity substantially increased on complexation with metals.


Precolumn derivatization of nucleotides based on fluorescent carbamate formation of the sugar moieties in high-performance liquid chromatography. The fluorescence derivatization of nucleotides with 2-(5-chlorocarbonyl-2-oxazolyl)-5,6-methylenedioxybenzo++  furan in the presence of sodium azide and the separation of the derivatives by high-performance liquid chromatography are described. The reagent reacts with 5'-terminal hydroxyl groups of nucleotides to produce the corresponding fluorescent carbamates. The derivatives of mono- and oligonucleotides are separated by chromatography on a reversed phase column (TSKgel ODS-80TM) and the derivatives of octa- and deca-nucleotides on a size exclusion column (TSKgel G3000SWXL). The detection limits (signal-to-noise ratio = 3) are 0.8-6.0 pmol on column. 5'Phosphorylated nucleotide also gives a fluorescent derivative after alkaline phosphatase-mediated dephosphorylation.


Heat induced conformational changes generate mitogenicity to splenocytes by sclerogen from Sclerotinia sclerotiorum IFO 9395. The fungal mitogen, sclerogen, obtained from sclerotia of Sclerotinia sclerotiorum IFO 9395 showed significant mitogenic activity to murine splenocytes after heat denaturation in relation to polymerization. To evaluate the conditions generating mitogenicity, we performed several chromatographic and spectral analyses. After heat denaturation of sclerogen, significant reduction of intrinsic fluorescence, significant changes on the ultraviolet absorption spectrum, significant changes on the circular dichroism spectrum, and an extreme change of the surface charge to anionic, were observed. These results strongly suggested that local as well as overall conformational changes of sclerogen associated with a high molecular mass and polyanionic charges are important in generating mitogenicity to murine splenocytes.


Mannan-coated liposome delivery of gadolinium-diethylenetriaminepentaacetic acid, a contrast agent for use in magnetic resonance imaging. Gadolinium-diethylenetriaminepentaacetic acid (Gd-DTPA), a paramagnetic contrast agent for use in magnetic resonance imaging (MRI) was bound to stearylamine and incorporated into the liposomal membranes (Gd-DTPA liposomes). In addition, the Gd-DTPA liposomes were coated with mannan (cholesterol-aminoethylcarbamylmethyl mannan), a polysaccharide, to obtain the mannan-coated liposomes. An in vitro MRI study showed that the Gd-DTPA liposomes produced a greater intensity of contrast than did the Gd-DTPA solution with a reduced T1 relaxation time. Intravenous injection of the Gd-DTPA liposomes containing 153Gd or liposomes containing 153Gd or 14C-DTPA to mice showed an accumulation of Gd-DTPA primarily in the liver and lung. When the mannan-coated liposomes were administered, an increased uptake of Gd-DTPA by these tissues was demonstrated. The mannan-coated liposomes may enhance contrast of the liver in MRI at a lower dose of Gd-DTPA.


Effect of oral pretreatment with antibiotics on the hydrolysis of salicylic acid-tyrosine and salicylic acid-methionine prodrugs in rabbit intestinal microorganisms. We examined the hydrolysis mechanism of salicylic acid-tyrosine (salicyl-tyrosine) and salicylic acid-methionine conjugate (salicyl-methionine) in rabbits by exploring their behavior following intraduodenal and intracecal administration (72 and 36 mumol/kg, respectively: salicylic acid equivalent). A large amount of salicyl-methionine was absorbed following intraduodenal administration of salicyl-methionine, without being metabolized to salicylic acid in the small intestinal mucosa. On the contrary, salicylic acid was detected in the blood following intraduodenal administration of salicyl-tyrosine, suggesting that salicyl-tyrosine was metabolized in the small intestinal mucosa. After oral pretreatment of rabbits with kanamycin sulfate (6 x 400 mg) or tinidazole (6 x 160 mg), the hydrolysis of salicyl-tyrosine and salicyl-methionine following intracecal administration was inhibited significantly, indicating that the intestinal microorganisms were responsible for the biotransformation of these prodrugs. Furthermore, in rabbits orally pretreated with both kanamycin sulfate and tinidazole, a significant inhibition of salicylic acid formation from salicyl-tyrosine and salicyl-methionine following intracecal administration was observed.


Synthesis of new fluorescent nucleosides, 3-beta-D-ribofuranosylpyrazolo (3,2-i) purine derivatives and their cytotoxic activities. The novel nucleosides 3-beta-D-ribofuranosylpyrazolo(3,2-i) purine (8) and their 9-substituted bromo, nitro and amino compounds (3, 6 and 11) have been prepared from a fully protected 3-beta-D-ribofuranosyl(3,2-i)purine-9-carboxyamide 1 by bromodeamidation (ipso bromination). Compounds 3, 8 and 11 exhibited antileukemic activity against mouse leukemia L5178Y cells in culture, while the 9-substituted nitro, ester and amide compounds (6, 12 and 13) showed no cytotoxicity.


Phagocytic cell function in recurrent endotoxemia in sheep. Sepsis syndrome in severely traumatized patients is supposedly due to a blockade of the phagocytic cell system--for example, the reticulo endothelial system (RES) and polymorph nuclear leukocytes (PMNL), which cause insufficient elimination of bacterial substances (e.g., endotoxin). In contrast we found in a previous study using a model of acute endotoxemia that RES clearance is enhanced, while PMNL function gave evidence for decompensation. In order to clarify whether or not our results were just a matter of single dose endotoxemia, we have investigated RES and PMNL function in a sheep model with recurrent endotoxemia. A sheep receiving endotoxin at a dose of 1 microgram/kg body weight every 12 hours was monitored over a 5-day period. RES clearance was calculated by the half-life of Tc99 phytate each day. The PMNL function was determined by measuring chemiluminescence (CL) of isolated PMNL and total blood. The half-life of Tc99 phytate decreased from 56 min to 44 min with endotoxin (P less than or equal to 0.01) the second day and further to 42 min (P less than or equal to 0.002) on the third day. Values then returned to baseline until the end of study. Zymosan induced and luminol enhanced CL response indicated an acute cellular exhaustion after the first endotoxin dose. Under subsequent endotoxin administration, a daily attenuation of the acute response was noted in combination with baseline opsonin capacity. Our results give evidence that also in recurrent endotoxemia RES function is characterized by a sufficient clearance, whereas the PMNL function remains decreased and therefore possibly is responsible for the well-known clinical post-traumatic septic complications.


A novel xanthine derivative counteracting in vivo tumor necrosis factor alpha toxicity in mice. The xanthine derivative A 802715 (1-(5-hydroxy-5-methyl)hexyl-3-methyl-7- propyl-xanthine, Hoechst AG) caused dose-dependent protection against lipopolysaccharide (LPS)-induced lethal shock in mice. In animals which had received the compound, the LPS-induced increase of serum tumor necrosis factor (TNF alpha) levels was not significantly affected. Protection against LPS-induced lethality was observed not only when A 802715 was given 1 hr before or simultaneously with LPS but also when administered 1 hr after LPS challenge. Administration of 200 mg/kg of the compound 1 hr before challenge also fully protected against lethal shock induced by intravenous administration of recombinant murine TNF alpha. It is concluded that A 802715 counteracts TNF alpha toxicity and that the drug bears the potential of therapeutic intervention in septic shock.


Pyrimidine-enhanced purine uptake in hepatocytes and improved survival from hypovolemic shock. To assess the effect of pyrimidines on the incorporation of purines into rat hepatocytes, monolayer preparations of hepatocytes were incubated with radiolabeled precursors and their uptake determined. The uptake of purine precursors (inosine monophosphate, inosine, and hypoxanthine) was 27.8 +/- 1.31 nmoles per well, per 30 min. In the presence of added pyrimidines (uridine monophosphate, uridine, and orotate), this increased by 22%. Further enhancement (45%) was observed when L-dihydroxyphenylalanine (L-DOPA) plus pyridoxal-5'-phosphate was added to the purine/pyrimidine incubation medium. The effect of increased uptake of purines on survival after hypovolemic shock was also studied. Rats were bled to a blood pressure of 40 mmHg for 105 min and then treated with return of shed blood plus saline with or without purines and pyrimidines. Survival at 24 hr was 43% (15/35) in control animals, 66% (8/12) (P = 0.276) when treated with inosine monophosphate (IMP) and aspartate (60 mumoles/kg each), and 83% (10/12) (P = 0.037) for IMP with aspartate and orotate treatment (60 mumoles/kg each). Thus, the addition of orotate enhanced survival possibly by promoting salvage or by uptake of purines. Nucleotide concentrations in the livers of animals after shock which had received IMP and orotate demonstrated a return of both ATP and energy charge to normal, further indicating the value of this treatment.


Kinin metabolism in the perfused ventilated rat lung. II: Influence of ventilation, perfusion, and perfusate composition variation on bradykinin metabolism in uninjured lung. Bradykinin metabolism by peptidases of the pulmonary endothelium has been investigated in the previously uninjured, ventilated, and asanguinously perfused rat lung. The influence of short-duration (up to 20 min) abnormal ventilation and perfusion conditions on bradykinin metabolism was assessed. Neither variation of the oxygen concentration (0 to 45%) nor omission of carbon dioxide in the ventilatory gas altered bradykinin metabolism significantly. Tidal volume variation did not alter bradykinin metabolism, and exclusion of one lung from the perfusion circuit reduced the capacity to degrade bradykinin proportionately. Acidification of the perfusion medium to pH 5 did not alter bradykinin metabolism. Acetylsalicylic acid in the perfusate protected the lung from an otherwise irreversible pressure increase associated with high-dose bradykinin perfusion. Endotoxin and hydrogen peroxide in the perfusate did not alter bradykinin metabolism. However, ammonia in the ventilatory gas caused immediate pulmonary edema, diminished lung capacity to metabolize bradykinin and altered the pattern of bradykinin metabolic products. The pulmonary endothelium itself, in the absence of blood, maintains its capacity to metabolize bradykinin under an extraordinary range of conditions.


Pentoxifylline treatment of sepsis in conscious Yucatan minipigs. Recent evidence suggests that pentoxifylline (PTX) may be useful in the treatment of sepsis. We examined effects of PTX in a conscious swine model of sepsis. Yucatan minipigs (20-30 kg) were anesthetized and instrumented with catheters in the vena cava, aortic arch, pulmonary artery (Swan-Ganz thermodilution catheter), and peritoneum. Twenty-four hours after surgery, sepsis was induced by intraperitoneal (ip) injection of Escherichia coli bacteria (2 x 10(10) cfu/kg). Nonseptic pigs received intraperitoneal saline (5 ml/kg). PTX treatment (3 mg/kg/hr, iv; 1 mg/ml in 0.9% saline) and maintenance fluid (5 ml/kg/hr, iv) were started with bacterial infusion. An additional 60 cc/kg 0.9% saline bolus was administered iv at 1 hr. Pigs were monitored before and 1, 2, 5, and 24 hr after bacterial injection. Intraperitoneal injection of bacteria led to significant reductions in blood pressure and cardiac output and elevations in pulmonary wedge pressure and pulmonary vascular resistance. These effects were attenuated by PTX treatment. All septic animals demonstrated elevated creatinine, blood urea nitrogen, circulating endotoxin (LPS), and tumor necrosis factor concentrations, reductions in white blood cell and platelet counts, and peritonitis. None of these responses was altered by PTX treatment. We conclude that PTX may prove to be a useful therapeutic tool in the early treatment of septic shock but is limited in the scope of its effects.


Beneficial effects of SK&F 105809, a novel cytokine-suppressive agent, in murine models of endotoxin shock. SK&F 105809 is a structurally novel dual inhibitor of lipoxygenase and cyclooxygenase-mediated arachidonic acid (AA) metabolism, which has demonstrated antiinflammatory activity in rodent models of inflammation. In addition, the active metabolite of this compound, SK&F 105561, has been shown in vitro to inhibit the production of the inflammatory cytokine interleukin-1 (IL-1) in human monocytes stimulated with lipopolysaccharide (LPS). We report here that in vitro SK&F 105561 also blocks the production of tumor necrosis factor (TNF) from human monocytes (IC50 0.8-3 microM). Furthermore, in a murine model of endotoxin shock in which animals are injected with LPS in combination with D-galactosamine (D-gal), SK&F 105809 (10, 30, and 100 mg/kg p.o.), delivered 30 min prior to LPS/D-gal, caused a dramatic reduction in serum TNF (40-90%) and protected the animals from the lethal effects of this treatment. Similar results were obtained in a second model of endotoxin shock in which mice were sensitized with Propionibacterium acnes 10 days prior to LPS injection. In this system 100-fold higher levels of serum TNF are elicited than with the LPS/D-gal model. Treatment with SK&F 105809 (30 and 100 mg/kg p.o.) delivered 30 min prior to LPS resulted in 90-100% inhibition of serum TNF. Protection from the lethal effects of LPS was observed at these doses in the P. acnes/LPS model.


Role of the L-arginine-nitric oxide pathway in the changes in cerebrovascular reactivity following hemorrhagic hypotension and retransfusion. We studied the involvement of the L-arginine-nitric oxide pathway in the changes in cerebrovascular reactivity following hemorrhagic hypotension and retransfusion. Feline middle cerebral arteries were prepared from control animals killed under anesthesia and from anesthetised animals subjected to hemorrhagic hypotension (stepwise bleeding to 90, 70, and 50 mmHg, maintained for 20 min at each level) followed by retransfusion (20 min). Two-mm-long vessel segments were suspended in organ chambers containing Krebs-Henseleit solution (37 degrees C, gassed with 95% O2-5% CO2) for isometric force measurements. Contractions to noradrenaline (norepinephrine), relaxations to acetylcholine, ATP, adenosine, and SIN-1, a nitric oxide donor compound, were compared in the vessels of the control and hemorrhage-subjected animals. Contractile responses to noradrenaline were significantly enhanced after hemorrhage, whereas relaxations to acetylcholine, ATP, and adenosine were significantly reduced. Relaxations to SIN-1, however, remained unchanged. L-Arginine did not cause relaxations in control vessels but relaxed the arteries after hemorrhage and retransfusion. To clarify the involvement of the L-arginine-nitric oxide pathway in these alterations, we studied the effect of exogenous application of L-arginine, the precursor of endothelium-derived relaxing factor (EDRF) and NG-nitro-L-arginine (NOLA), a competitive antagonist of the EDRF-producing enzyme on the vascular responses in vitro. Similar to the effect of hypotension and retransfusion, NOLA enhanced the noradrenaline-induced contractions and inhibited the acetylcholine-induced and purinoceptor-mediated relaxations in the control arteries. In the control vessels in vitro, L-arginine treatment did not modify any contractile or relaxant response. At the same time, in vitro L-arginine treatment inhibited the hemorrhagic hypotension-induced enhancement of the contractions to noradrenaline and restored the diminished relaxations to acetylcholine (but not to ATP or adenosine). In the vessels after hemorrhage, NOLA neither further enhanced the already markedly enhanced noradrenaline-induced contractions nor further inhibited the relaxations caused by ATP and adenosine. In the case of acetylcholine, however, NOLA caused a further inhibition of the relaxations. The effect of in vivo L-Arg infusion (30 mg/kg initial bolus and 10 mg/kg/min infusion) during hemorrhagic hypotension and retransfusion on the in vitro vascular reactivity was similar to that of the in vitro L-arginine treatment. The present study demonstrates that even short-lasting hemorrhagic hypotension combined with retransfusion markedly inhibits nitric oxide-mediated, agonist-induced endothelium-dependent cerebrovascular responses. In vivo or in vitro L-arginine treatment is able to restore normal responses to some agents.(ABSTRACT TRUNCATED AT 400 WORDS)


Effect of alpha-adrenergic blockage on cellular Ca2+ during endotoxic shock. The effect of alpha-adrenergic receptor antagonists, phentolamine, and prazosin on cytosolic Ca2+ concentration, [Ca2+]i, was studied in hepatocytes during endotoxic shock. Rats were given intravenous injections of endotoxin (20 mg/kg), phentolamine (3 mg/kg) plus endotoxin (20 mg/kg), or prazosin (5 mg/kg) plus endotoxin (20 mg/kg). They were sacrificed 5 hr later, at which time the endotoxin-injected rats showed signs of shock. Isolated hepatocytes were prepared and employed for the measurement of [Ca2+]i under basal and hormone-stimulated (1 and 10 microM epinephrine) conditions by means of Quin 2 fluorescence technique. The apparent basal level of [Ca2+]i in endotoxic rat hepatocytes (mean +/- SE: 482 +/- 31 nM) was significantly higher (P less than 0.05) than in phentolamine plus endotoxin (242 +/- 73) and prazosin plus endotoxin (240 +/- 43) groups. A significant increase in hepatocyte [Ca2+]i occurred with epinephrine in the phentolamine plus endotoxin and prazosin plus endotoxin groups, but not in the group receiving endotoxin alone. Endotoxic rats showed a mortality rate of 75%, whereas phentolamine plus endotoxin and prazosin plus endotoxin groups showed a mortality rate of 38% and 20% respectively. These data suggest that the protective effect of alpha-adrenergic receptor antagonists during endotoxic shock may be mediated, in part, by attenuating the entrance of Ca2+ into endotoxic liver cells.


Cardiovascular response in canine endotoxic shock: effect of ibuprofen pretreatment. Ibuprofen pretreatment increases arterial pressure and reduces mortality in endotoxic dogs. The increase in arterial pressure may be caused by increases in arterial resistance, arterial sphincter tone, or both. Thus it is not clear if ibuprofen pretreatment prevents the hemodynamic effects of endotoxemia or merely masks such effects by producing concomitant increases in arterial resistance. Accordingly, this study was performed to determine the effects of ibuprofen pretreatment on arterial pressure-flow relations and other measures of cardiovascular function in a canine model of endotoxic shock. In 19 pentobarbital-anesthetized and splenectomized, closed-chest dogs, biventricular stroke volumes were measured with electromagnetic flow probes, and intrathoracic vascular and pleural pressures were measured with catheters. Instantaneous venous return curves (see Pinsky MR, J Appl Physiol 56:765, 1984) were generated during positive-pressure ventilation, and steady-state arterial pressure-flow relations, left ventricular function, peripheral vascular compliance, oxygen consumption/oxygen delivery ratio, and arterial blood lactate levels were measured during two sequential volume loading and removal (20 ml/kg) sequences. All but two dogs received a bolus infusion of Escherichia coli endotoxin (2 mg/kg) between the two fluid challenge runs. Eleven of the 17 endotoxic dogs also received ibuprofen (15 mg/kg) immediately before the initial fluid challenge. Ibuprofen pretreatment abolished all hemodynamic effects of endotoxin, whereas in the untreated group endotoxin caused decreases in calculated arterial outflow pressure and in peripheral vascular capacitance. Oxygen consumption remained constant despite changes in oxygen delivery in the nonendotoxic dogs and in the ibuprofen-pretreated dogs, whereas oxygen consumption covaried with oxygen delivery in the endotoxic dogs not pretreated with ibuprofen. Arterial lactate levels were higher after endotoxin infusion (2.1 +/- 0.5 to 3.1 +/- 0.6 mmol/liter; P less than 0.05 pre- to postvolume) but were not different between treatment groups. These data suggest that ibuprofen alters many, but not all, of the hemodynamic effects of endotoxin infusion in the dog.


Malpractice in obstetrics: a contribution of cases in southern Italy. Looking ahead to the abolition of many intra-European frontiers the Authors offer this contribution to the discussion of the medico-legal aspects of gynecological activity in the United Europe of 1993. It is to this end that they present the data from a review of the material which has been the subject of litigation on the part specialists in Southern Italy, also commenting on the case series in the light of the Italian provisions in force up to date.


Retroperitoneal abscess and vena cava thrombosis following normal pregnancy and delivery. A 24 year old primigravida underwent a normal pregnancy and delivery, and developed a retroperitoneal abscess and thrombosis of the vena cava late in the puerperium. A transvaginal drainage of the abscess was performed and the thrombosis of the vena cava treated with Heparin and thrombectomy. Full recovery was obtained.


Eicosanoid production in human umbilical cord vessels: effect of 17 beta-estradiol. This study was designed to investigate the effects of 17 beta-estradiol on prostanoid formation from exogenous arachidonic acid in homogenates of human umbilical cord vessels. The veins produced more prostanoids than the arteries and predominantly 6-keto-PGF1 alpha. 17 beta-estradiol had no effect on the rate of production of prostanoids in either vessels. Thus, at least in our in vitro system, the regulation of the vascular tone by prostanoids seems not to be altered by the addition of 17 beta-estradiol.


Variations of lymphocyte sub-populations in vulvar condylomata during therapy with beta-interferon. Several experiences induced us to consider genital HPV infection as an expression of a local immunodeficiency. The aim of our study was to research the effect of immunotherapy on the lymphocyte subpopulations and Langerhans cells in vulvar condyloma. Twenty women with persistent vulvar condylomata, treated with 2,000,000 IU/die of beta-interferon for 15 days, were submitted to vulvar biopsy before and 2-5 months after medical treatment. The frozen sections obtained were assayed with the following monoclonal antibodies: OKT4 (T helper lymphocytes), OKT8 (T suppressor lymphocytes), OKB7 (B lymphocytes) and S-100 protein (Langerhans cells). Using a morphometric evaluation, the average number of both intraepithelial and stromal lymphocyte subsets and of the intraepithelial Langerhans cells was assessed. In all the biopsies preceding the medical treatment we found a low number of T helper lymphocytes both in the epithelium and stroma, with inversion of T4/T8 lymphocyte ratio and rare presence of Langerhans cells. In patients with a good therapeutic response (50-100% of condyloma reduction) we observed an increase in intraepithelial T4 lymphocytes and a decrease in both intraepithelial and stromal T8 lymphocytes. In cases with persistent disease after therapy, the histological pattern was similar to that observed in the first biopsy, with the exception of a significant increase in the average number of Langerhans cells. Our data correlate the clinical response to the immunotherapy with the histology of lymphocyte subsets in the vulvar condylomata. The increase in Langerhans cells observed in patients with negative response may be interpreted with a probable inability of these cells to promote the immune reaction.


Myometrial hypertrophy and uterine metropathy without apparent organic cause: rate or responsibility. Between 1971 and 1980 more than 6,000,000 hysterectomies were performed in the USA. Dysfunctional uterine hemorrhage with non tumoral uterus and hypertrophic characteristic has been one of the principal indications, without possibility of definition as a pathological entity with its own characteristics. With all these premises the Authors have attempted to see, by means of morphometric studies, the myocyte characteristics and the proportion and composition of the uterine wall and at the same time eventual hormone-dependence of this phenomenon. For this they turn to to determination of oestrogen and progesterone receptors.


Carcinoma of the vulva: critical analysis of survival and treatment of recurrences. Prognostic variables in vulvar cancer include: stage, lymph nodal status, tumour diameter, vascular space invasion, depth of invasion. Treatment must in the cost/benefit balance, be adequate to these prognostic variables and surgery is today the cornerstone in vulvar cancer management. In early stages it is possible to control the disease with personalized radical non-mutilant operations limiting the high incidence of complications of the past, without endangering the 5-year survival rate. The risk of relapses is related to some factors such as site of tumour growth, depth of tumoural infiltration, lymphnode involvement and vascular space involvement. Relapses appear earlier in the groin than in the primitive sites and cases with more than 3 effected nodes have a higher incidence of recurrences in any site. Survival in related to localization and time of relapses. The treatment of recurrence is personalized for different sites, almost always surgical, in local recurrences.


A comparative study of propofol and thiopental as induction agents for elective caesarean section. Twenty women for elective caesarean section received either propofol 2.3 mg/kgr or thiopental 4.4 mg/kgr for induction of general anaesthesia. Maintainance was similar for both groups. Mean arterial pressure and heart rate were recorded non-invasively before anaesthesia, during intubation, one and five minutes after intubation. There were no significant differences in haemodynamic response between the two groups. During intubation heart rate rose in both groups, but remained increased five minutes after tracheal intubation only in the thiopental treated women (p less than 0.05). There was no significant neonatal depression as assessed by Apgar Scores and blood gas analyses. Propofol appears to be a suitable alternative to thiopental as an induction agent for obstetric anaesthesia.


Plasma and amniotic fluid concentration of fibronectin during normal and diabetic pregnancy. Fibronectin is a plasma glycoprotein which is involved in coagulation, platelet function, tissue repair and the vascular endothelial basement membrane. To ascertain the influence of pregnancy on plasma concentrations of fibronectin, we qualified plasma concentrations of fibronectin in normal pregnant women during the first, second and third trimester; at the time of delivery; and on the third day post partum, using a radial immunodiffusion plate procedure. The concentrations of fibronectin found in these samples were compared with the concentration of fibronectin in 20 pregnancies complicated by diabetes. Mean plasma concentrations of fibronectin rose significantly through pregnancy and were significantly higher during delivery. A decrease in the concentrations was noticed on the third post partum day. An even more significant decrease of maternal plasma concentrations was noticed during cesarean section in normal pregnancies as compared to the concentrations found at the time of normal delivery. Of the diabetic group of women studied, higher concentrations of plasma fibronectin were found at the time of cesarean section than at the time of delivery. Maternal plasma concentrations of fibronectin were significantly greater than amniotic fluid and umbilical cord plasma concentrations.


Evidence for gene-environmental interactions in Utah families with hypertension, dyslipidaemia and early coronary heart disease. 1. Among 45,258 Utah families surveyed, about 4% have a strong aggregation of early coronary disease. In detailed clinical evaluation, about 21% of such high risk coronary families were found to have familial dyslipidaemic hypertension (FDH) and about 3% were found to have heterozygous familial hypercholesterolaemia (hFH). 2. Common and potentially modifiable environmental factors seem to play an important role in these high risk families. Non-genetic obesity promotes the expression of FDH. A high fat diet promotes the expression of FH. Cigarette smoking promotes earlier death in all coronary prone families. 3. Practical approaches are suggested for helping coronary prone pedigrees by applying our understanding of genetic and environmental factors that promote earlier coronary disease onset.


Membrane fluidity as a genetic marker of hypertension. 1. The purpose of the present study was to examine changes in membrane fluidity in hypertension by means of an electron spin resonance (ESR) and a spin labelling methods. 2. Erythrocytes from spontaneously hypertensive rats (SHR) and from patients with essential hypertension were examined and compared with those from age-matched normotensive controls. ESR spectra were obtained for a fatty acid spin label agent (5-nitroxide stearate) in the membranes. The values of outer hyperfine splitting (2T' parallel) and of order parameter (S) of the ESR spectra were significantly higher in erythrocytes from SHR and patients with essential hypertension than in those from normotensive controls. Similar results were obtained in cultured vascular smooth muscle cells of SHR. This finding shows that the membrane fluidity might be lower in SHR and in essential hypertension. 3. When Ca was loaded to erythrocytes with a Ca-ionophore (A23187), the parameters of the ESR spectra showed a greater increase (membrane fluidity was decreased) in SHR and in patients with essential hypertension than that in the normotensive controls. The Ca-induced alterations in membrane fluidity were not definitely observed in secondary hypertension. 4. These results suggest that the lower membrane fluidity might be a genetically determined abnormality of hypertension. The marked reduction of the membrane fluidity by Ca-loading in SHR and in essential hypertension might support the hypothesis that an abnormality of the Ca-handling at cellular levels could affect physical properties of the biomembranes in genetic hypertension.


Analysis of the apolipoprotein B3' hypervariable region in patients with essential hypertension. 1. The typing of the apolipoprotein B 3' hypervariable region was investigated in hypertensive and normotensive subjects using rapid typing of a variable number of tandemly repeated short DNA sequences (VNTR) by the polymerase chain reaction. 2. In the DNA samples of 89 normotensive and 99 hypertensive patients, 13 different-sized alleles were detected. The most frequent allele has 35 repeat units in both groups with frequencies of 0.624 and 0.596 in normotensive and hypertensive patients, respectively. Frequency distribution of 13 alleles was similar in both groups. 3. These results demonstrate no association between the apolipoprotein B gene polymorphism and essential hypertension.


Salt and hypertension: revisited.

Salt and hypertension: revisited. 1. There is only one known environmental factor that possibly could hold the key to much of the problem of 'essential' hypertension and this is salt. Without a high or moderately high salt intake, other environmental factors may be largely inoperative. The evidence, which still falls somewhat short of certainty, is briefly reviewed.


Experimental studies on cardiovascular disorders and cellular Ca handling in shell-less chick embryos. 1. Chick embryos rendered Ca deficient by long-term culture outside the eggshell develop hypertension/tachycardia. 2. In this study, the hypertension of the shell-less (SL) chick embryo has been characterized by comparing their cardiovascular response to adrenergic drugs and cellular Ca handling function with normal (NL) embryos. 3. Before and after the administration of noradrenaline (NA), phentolamine (PA), isoproterenol (IP), and propranolol (PP), blood pressure and pulse rate were measured. 4. Baseline blood pressure was consistently higher in SL embryos. In both embryos, blood pressure was elevated by NA and PP, but lowered by IP and PA. Pharmacological sensitivity to NA and PA was higher in SL embryos. Serum Ca was lower in SL embryos, while myocardial calcium content was not different. At the cellular level, Ca handling was studied using red blood cells (RBC) from SL and NL embryos under varying osmotic and ionic conditions by analysing 45Ca uptake and cell volume regulation. 5. Two features of RBC Ca uptake were apparent: (i) Na-Ca exchange was functional since Ca uptake was dependent on external Na; (ii) Ca uptake was dependent on total ionic electrochemical gradient. The SL RBC had greater ion permeability, more active Na-Ca exchange function and higher Ca pumping out capacity. 6. These systemic and cellular properties of the SL embryo indicate that it is a novel and useful experimental system to study the relationship between Ca homeostasis and the development of hypertension.


Analysis of factors in hypertension: change of membrane fluidity in the arteries of spontaneously hypertensive and monocrotaline-injected rats. 1. A change of membrane fluidity of a small piece of artery tissue was monitored by Fourier transform infra-red spectroscopy. 2. The measurement of carotid artery in situ revealed that a significant change in the peak position of the methylene absorbance was detected in the spontaneously hypertensive rat subjected to anoxia, but not in the Wistar-Kyoto rat carotid. The shift of a peak to the higher wavenumber position suggested that the averaged membrane fluidity of the artery was increased by the anoxic treatment. A similar change of membrane fluidity was also observed in a pulmonary hypertension induced by monocrotaline. 3. At 4 weeks after the injection of monocrotaline, the membrane fluidity was increased in the pulmonary artery tissue. 4. The change of membrane fluidity may be caused by the change of activity of phospholipases in the arteries with hypertension.


The role of oxidized lipoproteins in the pathogenesis of atherosclerosis. 1. Oxidized low density lipoprotein (LDL) has been suggested to play an important role in atherogenesis by facilitating the accumulation of lipids in macrophages. In vitro studies from this laboratory have shown that oxidized LDL is recognized not only by the specific receptor for it, but also by the receptor which is common for oxidized LDL and acetyl LDL. Probucol, originally developed as an antioxidant, prevents the oxidative modification of LDL in vitro. Recent studies by the authors show that probucol prevents the progression of atherosclerosis in homozygous Watanabe heritable hyperlipidaemic rabbits in vivo without any changes in plasma LDL cholesterol levels. 2. These results strongly suggest that oxidative modification of LDL could occur in vivo and probucol could slow the progression of atherosclerosis, without changes in plasma cholesterol levels. 3. In addition, recently the authors demonstrated that high density lipoprotein (HDL) particles are also oxidized. Once HDL particles were oxidized, they showed a lessened effect on the decrease of cholesteryl ester in foam cells, suggesting oxidative modification of HDL may stimulate development of atherosclerosis by limiting efflux of cholesterol from foam cells. Moreover, HDL particles from probucol-treated patients are hardly oxidized; subsequently, these HDL particles caused a marked efflux of cholesterol from foam cells.


Gene-environment interaction in hypertension, stroke and atherosclerosis in experimental models and supportive findings from a world-wide cross-sectional epidemiological survey: a WHO-cardiac study. 1. Genetic rat models of hypertension, such as spontaneously hypertensive rats (SHR) and in particular, stroke-prone SHR (SHRSP) are useful models for research on the genetic pathogenesis, gene-environment interaction and control of environmental factors for the prevention of hypertension and related cardiovascular diseases (CVD). 2. Since recent genetic analysis of hypertension in SHRSP indicated that one of the hypertensive genes related to salt-induced blood pressure (BP) rise was linked with the gene of the angiotensin-converting enzyme, gene-environment interaction is important in the humoral, neural, vascular and nutritional mechanisms of hypertension and CVD. 3. Extensive experimental studies in SHRSP by the authors have demonstrated nutritional factors counteracting directly or indirectly against the adverse effect of excess salt intake, such as K, Mg, Ca, dietary fibres, protein, some amino acids and fatty acids, etc.; they are therefore effective in preventing stroke, the typical complication of hypertension. 4. Experimental atherosclerotic models were established in SHRSP, which indicated also a gene-environment interaction in the pathogenesis of atherosclerosis at the cellular level of vascular smooth muscle cells. Excess salt intake accelerated cholesterol absorption from the intestine to induce arterial fat deposition as well as to active platelet aggregation by the mechanism of increased intracellular Ca2+ mobilization. 5. Based on such experimental findings on the hypertension and related CVD prevention, a cross-sectional multicentre epidemiological 'CVD and Alimentary Comparison' Study (WHO-CARDIAC Study) was designed to assess the relationship of biological markers of dietary factors with BP ('core' study) and major CVD mortalities ('complete' study) and has been successfully undertaken for the past 8 years in co-operation with 54 centres in 23 countries. 6. The results of 'core' study so far obtained by cross-centre simple linear regression analysis demonstrated adverse effects of body mass index and salt intake on BP in men and possible beneficial effects of Mg by within-centre multiple linear regression analysis. 7. Preliminary cross-centre simple linear regression analysis of the 'complete' study indicated a positive correlation of serum cholesterol levels and negative correlation of serum phospholipid poly-unsaturated to unsaturated fatty acid ratios and 24 h urinary taurine excretion with coronary heart diseases mortality; a positive correlation of 24 h urinary Na and Na/K ratios and a negative correlation of serum cholesterol levels with stroke mortalities was indicated. 8. These epidemiological data confirmed mostly the applicability of experimental findings to the nutritional prevention of hypertension and CVD prevention.(ABSTRACT TRUNCATED AT 400 WORDS)


Abnormal hsp70 gene expression: its potential key role in metabolic defects in hypertension. 1. Abnormal heat stress gene hsp70 expression has been demonstrated in organs as well as in cultured cells from spontaneously hypertensive rats and mice. 2. The polymorphism of hsp70, which is localized in the major histocompatibility complex, appears to segregate with a significant portion of hypertension. 3. Abnormal hsp70 expression may lead to anomalies in its feedback regulation, particularly since the steady state level of heat stress protein HSP70 seems to be lower in hypertensive than in normal animals. 4. Another biological consequence could be a modification of the effectiveness of cortisol, which may be involved in metabolic components of the pathogenesis of hypertension.


The association between salt sensitivity of blood pressure and family history of hypertension. 1. The salt sensitivity index (SSI) and family history of hypertension were studied in 140 hospitalized patients with essential hypertension to clarify whether salt sensitivity of blood pressure is related to familial disposition to hypertension. 2. SSI was calculated by dividing the change of mean blood pressure by that of urinary sodium excretion when salt intake was restricted from 15 to less than 3 g/day. 3. Family history of hypertension was classified into three groups depending on the presence or absence of hypertension in the father, mother and siblings. 4. The group without a family history of hypertension showed a significantly lower SSI value than other groups. 5. In multiple regression analysis undertaken within each gender, SSI showed significant partial correlations with blood pressure and family history of hypertension in the female group (r = 0.402 and 0.265, respectively), whereas in the male group it showed a positive correlation only with blood pressure (r = 0.501). These results indicate that salt sensitivity of blood pressure is related to familial disposition to hypertension. This association was more apparent in the female than male group and its gender difference can be partially attributed to the fact that blood pressure in the female group is more sensitive to salt.