Selective binding to AT sequences in DNA by an acridine-linked peptide containing the SPKK motif. The sequence selectivity of binding to DNA by an acridine-linked peptide ligand has been investigated by means of footprinting methodologies. The ligand conjugates an anilino-acridine intercalating chromophore with the potentially minor groove binder octapeptide SPKKSPKK. This basic peptide corresponds to a highly conserved DNA recognition motif found in histone H1 and several other nonhistone proteins. Three complementary techniques using DNase I, hydroxyl radicals and osmium tetroxide as sequencing probes have been employed to evaluate both the sequence specificity of binding and the drug-induced conformational changes in DNA. The results converge to demonstrate the AT-selectivity and support a model in which the peptide moiety lies in the minor groove. DNA-binding sites of the conjugate are restricted to a few alternating AT-sequences proximal to GC-rich regions. Binding to homooligomeric runs of A and T is clearly disfavoured by the hybrid whereas such sequences represent preferred binding sites for the unsubstituted basic peptide. These differences reflect the influence of the anilino-acridine chromophore, which evidently contributes to the DNA recognition process allowing the peptide only to contact defined DNA sequences.


A potent and specific agonist, Suc-[Glu9,Ala11,15]-endothelin-1(8-21), IRL 1620, for the ETB receptor. A series of C-terminal linear peptides of endothelin (ET)-1 and their N alpha-succinyl (Suc) analogs were synthesized and their binding affinities for the two subtypes of ET receptor, ETA and ETB, in porcine lung membranes were examined. Among the synthetic analogs, Suc-[Glu9,Ala11,15]-ET-1(8-21), IRL 1620, was the most potent and specific ligand for the ETB receptor (KiETA/KiETB approximately equal to 120,000) as judged by the Ki values for ETA (1.9 microM) and ETB (16 pM) receptors. IRL 1620 was 60 times more selective for the ETB receptor than ET-3 (KiETA/KiETB approximately equal to 1,900). IRL 1620 (10(-9)-10(-7) M) induced contractions of the guinea pig trachea with a comparable potency to those of ET-1 or ET-3, suggesting that IRL 1620 is a potent ETB receptor agonist.


Cyclic inhibition-potentiation of the crosslinking of synapsin I with brain microtubules by protein kinase FA (an activator of ATP.Mg-dependent protein phosphatase). The ATP.Mg-dependent type-1 protein phosphatase activating factor (FA) was identified as a protein kinase that could phosphorylate synapsin I, a neuronal protein that coats synaptic vesicles, binds to cytoskeleton and is believed to be involved in the modulation of neurotransmission. More importantly, more than 90% of the phosphates in 32P-synapsin I phosphorylated by FA could be removed by the activated ATP.Mg-dependent type-1 protein phosphatase and the synapsin I phosphatase activity was found to be strictly FA-dependent. Functional study further revealed that as a synapsin I kinase, factor FA could phosphorylate synapsin I and thereby inhibits crosslinking of synapsin I with tubulin, while as a synapsin I phosphatase activator, FA could promote the crosslinking copolymerization of synapsin I with tubulin. Taken together, the results provide initial evidence that a cyclic modulation of the crosslinking copolymerization of synapsin I with brain microtubules can be controlled by factor FA, representing an efficient cyclic cascade control mechanism for the regulation of axonal transport process during neurotransmission.


Immunochemical characterization of a cytochrome P450 isozyme and a protein purified from liver microsomes of male guinea pigs and their roles in the oxidative metabolism of delta 9-tetrahydrocannabinol by guinea pig liver microsomes. A protein (designated as protein-B) was purified from liver microsomes of adult male guinea pigs by an affinity chromatography with omega-aminooctyl Sepharose 4B, followed by HPLC using DEAE-5PW and hydroxyapatite columns which had been used to purify a cytochrome P450 (P450) isozyme (P450-A) from the same subcellular fraction (Narimatsu et al., Biochem Biophys Res Commun 172: 607-613, 1990). Protein-B had a molecular mass of 49 kDa in SDS-PAGE, but did not show absorbance at 417 nm for heme. Further, it did not show any oxidative activities towards aniline (AN), d-benzphetamine (d-BP), p-nitroanisole (p-NA) or delta 9-tetrahydrocannabinol (delta 9-THC) in a reconstituted system including dilauroylphosphatidylcholine, NADPH-P450 reductase, and cytochrome b5. However, antiserum against protein-B raised in rabbits suppressed liver microsomal oxidative activities towards d-BP and p-NA dose-dependently. The antibody decreased delta 9-THC oxidative activity most effectively, but did not decrease AN hydroxylation activity. Antiserum against P450-A suppressed all the activities towards these four substrates, especially towards delta 9-THC, in liver microsomes of male guinea pigs. Moreover, reconstitution with hemin made it possible for protein-B to produce some oxidative activity toward delta 9-THC. These results suggest that protein-B is also a cytochrome P450 isozyme which has lost a heme moiety during purification steps. Both P450-A and protein-B could have a role as cytochrome P450 isozymes in the oxidative metabolism of drugs, especially that of delta 9-THC by the liver microsomes of adult male guinea pigs.


Characterization of the thromboxane receptor mediating prostacyclin release from cultured endothelial cells. The thromboxane A2 (TXA2) mimetic, 9,11-dideoxy-11,9-epoxymethano-prostaglandin F 2 alpha (U46619), mobilized calcium in the bovine aortic endothelial cell line AG4762 and stimulated release of prostacyclin from these cells. The U46619-stimulated release of prostacyclin could be inhibited by TXA2 antagonists with the order of potency [Is-[1 less than a, 2 less than b(5z), 3 less than b, 4 less than a]]-7-[3-[[2-[(phenylamino)carbonyl]hydrazino]methyl]-7-oxabicyclo- [2.2.1]hept-2-yl]-5- heptenoic acid (SQ29548) greater than 4-[2-(4-chlorobenzene-sulphonamido) ethyl]phenylacetic acid (BM13505) greater than 4-[2-(phenylsulphonamido)-ethyl]phenoxyacetic acid (BM13177), which was consistent with release being mediated by a TXA2 (TP) receptor. The TP receptor ligands, [3H]SQ29548 and 9,11-dimethylmethano-16(3-[125I]iodo-4-hydroxyphenyl)-13,14-dih ydr o-13-aza- 15-omega-o-tetranor-thromboxane ([125I]-PTA-OH), both appeared to bind to a homogenous population of sites in AG4762 cell membranes. The affinities of [3H]SQ29548 and [125I]PTA-OH were approximately 10 nM and approximately 0.3 nM, respectively, and the density of sites labelled by either ligand was approximately 25 fmol/mg protein. Under conditions where equilibrium was approached, the specific binding of [3H] SQ29548 or [125I]PTA-OH was displaced by SQ29548, BM13505 and BM13177 with the same order of potency and similar apparent affinities as in the functional assay, suggesting that these binding sites represent bona fide TP receptors.


Reactions of glutathione with the catechol, the ortho-quinone and the semi-quinone free radical of etoposide. Consequences for DNA inactivation. Etoposide [4'-demethylepipodophyllotoxin-9-(4,6-O-ethylidene-beta- D-glucopyranoside)] can be metabolized to DNA-inactivating catechol, ortho-quinone and semi-quinone free radical derivatives which may contribute to its cytotoxicity. In this paper, we examined in vitro whether glutathione (GSH), which is known to react easily with quinoid compounds, could interact with the active etoposide intermediates and in this way influence the cytotoxicity of the parent compound. To this end, reactions of GSH with the etoposide intermediates were studied, using HPLC and ESR measurements, together with the effects of GSH on the biological inactivation of single-stranded (ss) and double-stranded (RF) phi X174 DNA by these compounds. From the results it could be determined that: (a) GSH does not react with the catechol and, as a consequence, has no effect on the reaction of this intermediate of etoposide with ss and RF phi X174 DNA; (b) GSH reacts with the ortho-quinone most likely by formation of a conjugate and by two-electron reduction to the catechol, resulting in a partial protection of ss and RF phi X174 DNA against inactivation by this species; and (c) GSH protects ss phi X174 DNA against inactivation by the semi-quinone free radical of etoposide probably by conjugation with this species.


Specific inhibition of calcineurin by type II synthetic pyrethroid insecticides. The inhibitory action of synthetic pyrethroids and some chlorinated hydrocarbon insecticides on the neural calcium-calmodulin-dependent protein phosphatase, calcineurin, was studied using one radiotracer and two colorimetric methods. It was found that all insecticidal Type II pyrethroids (cypermethrin, deltamethrin and fenvalerate) are potent inhibitors of isolated calcineurin from bovine brain. Their IC50 values were approximately 10(-9) to 10(-11) M. By contrast, neither noninsecticidal chiral isomers of these pyrethroids, neuroactive Type I pyrethroids nor neuroactive chlorinated hydrocarbon insecticides showed comparable potencies against this enzyme. To confirm the action of Type II pyrethroid in situ, isolated intact rat brain synaptosomes were incubated with [32P]phosphoric acid and subsequently depolarized in the presence and absence of 0.1 microM deltamethrin. As expected, there was a sharp rise in protein phosphorylation due to the action of calcineurin. Deltamethrin caused a distinct delay in the dephosphorylation process. The results clearly indicate that calcineurin is specifically inhibited by Type II pyrethroids.


Beta-adrenergic responsiveness in cultured aorta smooth muscle cells. Effects of subculture and aging. beta-Adrenoceptor-mediated vasorelaxation is diminished in vessels from a variety of aged species including humans. This phenomenon was studied for the first time in cultured aorta smooth muscle cells (ASMC) from young (4- to 6-month) and old (24- to 26-month) F-344 rats. Cyclic AMP (cAMP) accumulation was assessed following isoproterenol and forskolin stimulations in primary cultures and after 1-4 passages of aorta smooth muscle cells. Isoproterenol and forskolin increased cAMP accumulation 6- and 10-fold, respectively, in primary cultures from young rats. Isoproterenol stimulation was reduced markedly in passaged cells. Forskolin stimulation was unaffected, indicating passage-related phenotypic changes in receptor-mediated stimulation, but not in post-receptor adenylate cyclase activation. The response to isoproterenol was diminished in old animals, but that to forskolin was unaltered. Thus, cultured ASMC from F-344 rats are highly responsive to beta-adrenoceptor stimulation and demonstrate age-related changes, but undergo phenotypic modulation during passage.


Characterization of solubilized bradykinin B2 receptors from smooth muscle and mucosa of guinea pig ileum. Bradykinin (BK) B2 receptors in guinea pig ileum were characterized in both membrane and soluble form. [3H]BK bound to a single class of sites with almost identical affinities in membranes prepared from the longitudinal muscle, circular muscle and mucosal layers of the ileum. The pharmacology of the binding in the distinct layers was indistinguishable. The detergent 3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulfonate (CHAPS) maximally solubilized nearly 80% of membrane binding activity in a very stable conformation. In soluble preparations, [3H]BK labeled a single class of sites but with about 10-fold lower affinity. The affinities of BK analogs in competition studies were similarly reduced. There was no difference in the pharmacology of the binding in soluble receptors prepared from the different layers of the ileum. The results show that the ileum is a good source of solubilized B2 receptors and that the receptors in the smooth muscle and the mucosa are very similar.


Beta-naphthylamine induces anion superoxide production in rat peritoneal macrophages. Rat peritoneal macrophages were incubated in the presence of beta-naphthylamine (beta-NA), a well known carcinogenic agent, and some parameters of respiratory burst were studied. beta-NA induced a time- and dose-dependent stimulation of superoxide anion (O-2) production, and this enhancement was suppressed by the addition of superoxide dismutase enzyme. Also, no cooperative effect between beta-NA and phorbol 12-myristate 13-acetate was observed. Other observations were as follows: (i) the simultaneous presence of polymyxin B, and staurosporine inhibitors of protein kinase C, inhibited beta-NA-dependent O-2 production; (ii) NADPH-oxidase contained in postnuclear fraction from beta-NA-incubated macrophages showed a greater activity than control fractions; (iii) the stimulation of O-2 production elicited by beta-NA was several-fold enhanced in activated macrophages compared to resident cells. These data suggest that beta-NA produces the activation of NADPH-oxidase through protein kinase C.


Lymphocyte subsets associated with T cell receptor beta-chain gene rearrangement in patients with rheumatoid arthritis and neutropenia. To determine the incidence of a clonal lymphoid disease in patients with chronic rheumatoid arthritis (RA) and neutropenia. Lymphocytes from 23 RA patients with either current neutropenia or a history of this complication were studied. Eight patients had a clonal rearrangement of the T cell receptor beta-chain gene. Phenotypically, they showed a distinctive pattern characterized by an inverted CD4+:CD8+ cell ratio and an increased number and percentage of CD57+/CD8+ and CD3+/DR+ lymphocytes. None had evidence of a lymphoid malignancy. Among RA patients with neutropenia, there is a subset who have a subclinical disease resembling T gamma lymphoproliferative disease.


The effects of low density lipoprotein and high density lipoprotein on phosphoinositide hydrolysis in bovine aortic endothelial cells. Low density lipoprotein (LDL) and high density lipoprotein (HDL3) were tested for their ability to induce inositol phospholipid turnover and inositol phosphate production in bovine aortic endothelial cells (BAEC). The production of inositol phosphates following hydrolysis of the phosphoinositides was demonstrated by two methods; release of [3H]inositol phosphates after labelling with [3H]myo-inositol and by a direct binding assay for inositol 1,4,5-trisphosphate (InsP3). Acute exposure to LDL induced InsP3 release at low concentrations of the lipoprotein within the physiological range of LDL in tissues. HDL3 did not cause any release of the inositol phosphates. Pre-incubation of BAEC with HDL3 suppressed bradykinin- and LDL-induced inositol phosphate production in BAEC in a concentration-dependent manner. It is concluded that LDL acutely stimulates phosphoinositide breakdown and that pre-incubation of cells with HDL3 inhibits this effect. The mechanism responsible for these effects remains to be elucidated.


[Methods of risk assessment from data of experimental carcinogenesis studies]. A comparison of the carcinogenic potencies of different substances and an assessment of the risk to exposed people can contribute significantly to the management of carcinogenic hazards including adequate regulatory decisions. A mathematical estimation of the risk caused by a certain height of exposure is usually done under the assumption of a linear dose-response relationship by means of an arithmetical factor which gives the ratio of risk to dose in the low-response range and which has to be found on the basis of epidemiological or experimental data. This work deals with calculations of risk/dose ratios from the data of three carcinogenicity tests by means of statistical methods. The influence of the selection of mathematical model, background assumption and the point of the function chosen for the calculation of the ratio was tested. The results show that an additivity assumption for the background leads to linearity of the curve in the low-response range such that the ratios calculated with different models (multistage, weibull, logit, probit) or for different points of the functions do not differ significantly within the data sets tested. Even under the assumption of independence of the background risk the influence of the model selected is still small if the ratios are calculated for the lowest experimental dose or for an exposure associated risk of 1%. Risk/dose ratios calculated by one of these methods utilize the information of several experimental groups. They seem to be well suited for a direct comparison of the potency of different carcinogens and as a basis for an assessment of the carcinogenic risk to humans, which can serve as a measure of the hazard of a single exposed person or may give an idea about the tumour incidences to be expected within an exposed population.


[The detection of infectious rotaviruses by cell culture technique: use and evaluation of the immunoperoxidase assay]. In the present study, virus isolation was attempted in one hundred and twenty-one fecal samples of children suffering from acute gastroenteritis. Virus isolation was performed either conventionally by examination of cytopathogenic effect (CPE) or by immunoperoxidase staining (IPS) of rotavirus group specific antigen (inner capsid) in trypsin free MA104 cells within 18 h. Applying the conventional technique, rotavirus was isolated in only 4 (3.3%) fecal specimens. In contrast, IPS detected infectious virus in 49 (40.5%) stool samples. The specificity of IPS was confirmed by the results obtained with an antigen detection ELISA ("Rotazyme", Abbott, Wiesbaden) and gel electrophoresis of rotaviral RNA (electrophoretyping). ELISA and RNA gel electrophoresis detected rotavirus in 66 (54.4%) and 56 (46.3%) stool samples respectively. IPS enables rapid diagnosis of rotavirus infections in cell cultures applied to detect infectious viral particles in order to be used in the investigation of nosocomial outbreaks, material- and surface contamination and evaluation of disinfectants.


[Bacteriologic and virologic hygienic studies on waste water ponds]. Many problems arise with the overflow of wastewater after heavy rainfalls. Therefore a research project was started to investigate the possibilities of biological treatment for such sporadically occurring sewage. In this paper a helophyte bed is compared with a simple wastewater pond regarding virological and bacteriological hygiene parameters. The number of coliphages is an evident indicator for enteroviruses and its enumeration can be done easily and safely. It is suggested that the kind of sewage treatment described here is not suitable for municipal wastewater but for domestic sewage in the countryside. Furthermore the quantification of coliphages should become a constant part of routine hygiene examination.


Glucose absorption, hormonal release and hepatic metabolism after guar gum ingestion. Six non-anaesthetized Large White pigs (mean body weight 59 +/- 1.7 kg) were fitted with permanent catheters in the portal vein, the brachiocephalic artery and the right hepatic vein and with electromagnetic flow probes around the portal vein and the hepatic artery. The animals were provided a basal none-fibre diet (diet A) alone or together with 6% guar gum (diet B) or 15% purified cellulose (diet C). The diets were given for 1 week and according to a replicated 3 x 3 latin-square design. On the last day of each adaptation period test meals of 800 g were given prior to blood sampling. The sampling was continued for 8 h. Guar gum strongly reduced the glucose absorption as well as the insulin, gastric inhibitory polypeptide (GIP) and insulin-like growth factor-1 (IGF-1) production. However, the reduction in peripheral blood insulin levels caused by guar gum was not associated with a change in hepatic insulin extraction. IGF-1 appeared to be strongly produced by the gut. The liver had a net uptake of the peptide. Ingestion of guar gum increased the hepatic extraction coefficient of gut produced IGF-1. Guar gum ingestion also appeared to decrease pancreatic glucagon secretion. Cellulose at the level consumed had very little effect on the parameters considered. It is suggested that the modulation of intestinal mechanisms by guar gum was sufficient to mediate the latter internal metabolic effects.


Heterologous supersensitization between serotonin2 and alpha 2-adrenergic receptor-mediated intracellular calcium mobilization in human platelets. Recent reports suggest that serotonin (5-HT)2 receptor-mediated second messenger systems are enhanced in platelets of affective disorders. To make the mechanism of the enhanced response clear, we investigated 5-HT2 and alpha (alpha) 2-adrenergic receptor-induced intracellular calcium (Ca2+) mobilization in platelets of healthy volunteers, using fura-2. 5-HT2 and alpha 2-adrenergic receptor-mediated Ca2+ mobilization was enhanced by prior exposure to the other type of agonist, so called "heterologous supersensitization." The supersensitization was due to the enhancement of maximal response without change in agonist affinity. Chelating extracellular Ca2+ did not diminish the supersensitization. This enhancement of Ca2+ mobilization was not inhibited by H-7, an inhibitor of protein kinase C. However, this supersensitization was inhibited by pretreatment with sodium fluoride which directly activates guanine nucleotide binding regulatory proteins (G proteins). These results suggest that the supersensitization was caused from intracellular Ca2+ storage sites through a G protein-coupled pathway.


Selective synthetic ligands for human nuclear retinoic acid receptors. From a series of naphthalene and benzoic acid derivatives we have identified synthetic retinoic acid analogues exhibiting high selectivity for the nuclear retinoic acid receptors RAR alpha (Am 580), RAR beta (CD 2019) and RAR gamma (CD 437) as well as ligands sharing high affinities for all RAR subtypes (CD 367). The compounds were evaluated in two complementary screening systems: (1) binding to nuclear proteins extracted from COS-7 cells after transfection with the appropriate expression vectors, and (2) induction of plasminogen activator in the embryonic mouse teratocarcinoma cell line F9. All compounds behaved as retinoic acid agonists in the F9 test.


Detection of porin antigen in serum for early diagnosis of mouse infections with Salmonella typhimurium. The monoclonal antibodies to porin, an outer membrane protein isolated from Salmonella typhimurium and sandwich enzyme linked immunosorbent assay (ELISA) has made possible the detection of porin from sera of S. typhimurium-infected mice. The specificity of the monoclonal antibodies was ascertained based on their cross-reactivity with porins isolated from S. typhi, Shigella flexneri and Escherichia coli and lipopolysaccharide (LPS) of S. typhimurium and E. coli. Serum samples were found to be positive for porin as early as 3 days after intravenous and 5 days after oral infection. In addition, a positive correlation was observed between the bacterial load and the concentration of porin detected in the sera. On the other hand, analysis of sera for anti-porin antibody showed diametrically opposite time kinetics with antigenaemia. These results indicate that porin accumulates in the serum of infected mice much earlier than the appearance of antibodies to porin. Thus detection of porin holds promise for early diagnosis of typhoid.


Calcium channel blockers.

Calcium channel blockers. Calcium channel blockers are widely used in the treatment of ischemic heart disease, hypertension, and supraventricular tachycardia. The prototype agents, verapamil, nifedipine, and diltiazem, represent three classes of calcium channel blockers, each of which has different pharmacologic effects. Nifedipine and the other dihydropyridines primarily are vasodilators and have no clinical effects on cardiac conduction or contractility. Diltiazem and verapamil also are vasodilators, but they possess, to varying degrees, negative inotropic, chronotropic, and dromotropic effects. Side effects of these drugs are relatively rare and usually not serious, with the exception of potential conduction disturbances and heart failure in patients with underlying cardiac disease. To assess patients taking these medications and provide the necessary teaching, the nurse needs an understanding of the pharmacologic properties, clinical indications, and potential adverse effects of the various drugs.


Investigation of the biochemical effects of renin inhibition in normal volunteers treated by an ACE inhibitor. 1. In order to investigate accurately the biochemical effects of renin inhibition in man, we have developed a sensitive assay to measure angiotensin I (1-10) decapeptide. 2. Angiotensins were extracted from plasma by adsorption to phenylsilylsilica, and angiotensin I (Ang I) was quantified by radioimmunoassay. The detection limit was 0.77 fmol ml-1, and the extraction recovery of [125I]-Ang I added to albumin buffer was 83% at the inflection point (10 fmol ml-1) of the standard curve. The overall recovery was 98.5 +/- 3.5%. The intra- and inter-assay reproducibility was 10.4% and 9.7% respectively. Cross-reactivity of the antiserum used was low (less than 0.3%) with all angiotensin peptides tested except Ang (2-10) nonapeptide. 3. A human pharmacological model was subsequently used to assess in vivo the biochemical effects of the renin inhibitor CGP 38560A. Six healthy volunteers received 20 mg lisinopril, a long-acting ACE-inhibitor. During the following 24 h, the renin-angiotensin system was reset with typically elevated active plasma renin and Ang I, at respectively 275 and 429% of basal values. 4. In a randomized three-way cross-over protocol, the six volunteers received a 30 min infusion of the renin inhibitor CGP 38560A (125 or 250 micrograms kg-1) or 5% glucose. The fall in plasma Ang I was 92% and 97.5% after the lowest and highest dose of the renin inhibitor, respectively. A concomitant increase in active plasma renin was observed.(ABSTRACT TRUNCATED AT 250 WORDS)


Lack of effect of chronic calcium antagonist treatment on beta 1- and beta 2-adrenoceptors in right atria from patients with or without heart failure. 1. We studied the effects of chronic calcium antagonist (calcium entry blocker, CEB; nifedipine, verapamil, diltiazem) treatment on beta-adrenoceptor density (assessed by (-)-[125I]-iodocyanopindolol [ICYP] binding) and subtype distribution in right atria from 65 patients without apparent heart failure undergoing elective coronary artery bypass grafting (CAD-patients) and from 13 patients with moderate heart failure (NYHA class III to class III-IV) undergoing mitral valve replacement (MVD-patients). 2. In CAD-patients atrial beta-adrenoceptor density was 79.3 +/- 7.9 fmol ICYP bound mg-1 protein (n = 18), the beta 1:beta 2-adrenoceptor ratio 69:31%. Chronic CEB-treatment did not affect either atrial beta-adrenoceptor density or beta 1:beta 2-adrenoceptor ratio. 3. In contrast, in CAD-patients chronically treated with beta 1-adrenoceptor antagonists (atenolol, bisoprolol, metoprolol) and CEB, atrial beta-adrenoceptor density was significantly increased (108.6 +/- 10.5 fmol ICYP bound mg-1 protein, n = 21); this increase was due to a selective increase in beta 1-adrenoceptors. 4. In MVD-patients atrial beta-adrenoceptor density (55.5 +/- 8.7 fmol ICYP bound mg-1 protein, n = 7) was significantly lower (P less than 0.05) than in CAD-patients; beta 1:beta 2-adrenoceptor ratio, however, was not changed (67:33%). Chronic CEB-treatment of MVD-patients did not prevent the decrease in atrial beta-adrenoceptors. 5. We conclude that chronic CEB-treatment does not affect human right atrial beta-adrenoceptor density, either in patients without apparent heart failure or in patients with moderate heart failure.(ABSTRACT TRUNCATED AT 250 WORDS)


Sumatriptan and cerebral perfusion in healthy volunteers. 1. The effect of sumatriptan on regional cerebral perfusion was studied in healthy volunteers. 2. Intravenous sumatriptan (2 mg) had no detectable effect on regional cerebral perfusion as measured using a SPECT system with 99technetiumm labelled hexemethylpropyleneamineoxime. 3. Sumatriptan had no effect on pulse, blood pressure or ECG indices. 4. All six volunteers experienced minor adverse effects during the intravenous infusion.


The effect of orthotopic transplantation on total, beta 1- and beta 2-adrenoceptors in the human heart. 1. [125I]-(-)pindolol binding was used to determine beta-adrenoceptor density in homogenate preparations of right ventricular endomyocardial biopsies from 43 non-rejecting patients over the first 13 months following cardiac transplantation. The selective beta 1 subtype antagonist ligand CGP 20712A was used to determine the subtype density in 32 of the patients. Biopsy specimens from 15 donor hearts were used as controls. 2. beta-adrenoceptor density (expressed in terms of fmol mg-1 protein) was increased in the group of transplanted hearts as a whole compared with the donor hearts with respect to total (35 +/- 2 vs 23 +/- 2) and the beta 1 subtype (25 +/- 2 vs 16 +/- 2) whereas the beta 2 subtype and radioligand dissociation constant did not differ. 3. Non-parametric analysis of variance of total receptor density over time revealed significant heterogeneity which appears to be due to a discrete increase in beta-adrenoceptor density during the 4th post operative month. 4. These results indicate that beta-adrenoceptor density is not constant following transplantation. Furthermore, the increase in receptor density following transplantation is due mainly to an increase in the beta 1 subtype without a significant change in the beta 2 subtype.


The effect of respiratory manoeuvres and pharmacological agents on the pharmacokinetics of nedocromil sodium after inhalation. 1. Eight healthy subjects inhaled nedocromil sodium from a metered-dose inhaler using a standardised inspiratory technique. Blood samples were taken for up to 270 min after inhalation for radioimmunoassay of plasma nedocromil sodium concentrations. 2. To investigate the possibility that respiratory manoeuvres can alter the absorption of the drug from the lungs, on the first (control) study day at 70 min after dosing, subjects performed nine forced expiratory manoeuvres over a 3 min period. At 110 min after dosing, subjects took a slow, full inspiration with a 30 s breath-hold, and at 150 min after dosing the subjects performed one single forced expiration. 3. On the second study day, subjects inhaled methoxamine, 0.15 mg kg-1 of a 20 mg ml-1 solution at 60 min after dosing, and the study continued as above. On the third day, subjects repeated the sequence of respiratory manoeuvres, after having taken phenoxymethyl penicillin and probenecid by mouth for 48 h. 4. Both multiple forced expirations and the deep inspiration with breath-hold produced significant increases in the absorption of nedocromil sodium. Inhaled methoxamine did not alter airway calibre or the response to the respiratory manoeuvres. Probenecid, but not penicillin, was detected in the subjects' plasma, and had the effect of increasing the rise in plasma nedocromil sodium concentrations after the multiple forced expirations when compared with the control day. 5. These data suggest that disruption of epithelial tight junctions induced by the respiratory manoeuvres leads to enhanced paracellular transport of nedocromil sodium into the draining circulation of the airways and alveoli.(ABSTRACT TRUNCATED AT 250 WORDS)


Infection of Chang cells with hepatitis C virus using hepatic biopsy specimens from patients with chronic hepatitis (type C). The presence of hepatitis C virus sequence was detected in liver tissue extracts by the polymerase chain reaction (PCR) method using primers of non-coding region in six out of eight cases with chronic hepatitis seropositive for Chiron's antibody. Subsequently, liver extracts from these cases were added to cell cultures of Chang cells for 3 days. The liver extracts of the six cases positive for PCR appeared to infect the Chang cells.


The interaction of platinum antitumour drugs with mouse liver mitochondria. A study was undertaken to determine if cis-DDP and its second generation derivatives produced effects on mouse liver mitochondria, and if any of the observed effects could be correlated with the nephrotoxicity of the drugs. Although changes were observed in mitochondrial morphology, enzyme activity, Ca2+ influx, terbium binding and surface potential, no specific effect was correlated with nephrotoxicity. cis-DDP produced marked changes in mitochondrial morphology; electron probe analysis showed binding of the drug to the mitochondria. Inhibition of complex I and II activity of the respiratory chain and an ionic-strength-dependent effect on Tb3+ (a Ca2+ analogue) fluorescence were observed. The non-nephrotoxic derivatives, CHIP and tetraplatin, also produced significant changes in morphology. Treatment with these derivatives also produced decreases in mitochondrial enzyme activity, but the effect on terbium binding had an ionic-strength dependence which was inverse to that observed with cis-DDP. The tetravalent compounds also had a notable effect on mitochondrial surface potential. Carboplatin had an effect on morphology and Ca2+ influx and it inhibited the respiratory enzymes, although in a manner different from that observed with cis-DDP. Carboplatin had a minimal effect on terbium binding. It is evident that if the platinum drugs enter a cell to exert their action at the nuclear level, they will also depress mitochondrial function. The observed effects did not correlate with nephrotoxicity but, since all four compounds significantly altered mitochondrial structure and function, they may be related to the cytotoxicity of the drug.


Classification of calcium channels and calcium antagonists: progress report. The molecular biology and electrophysiology of calcium channels is proceeding apace, partially driven by the clinical success of the drugs classed as "calcium antagonists." Indeed, there are now more than 70 drugs in development that have been claimed to be "calcium antagonists" of one type or another. In order to ensure that there is a logical nomenclature for the channels and the drugs acting at the channels, an international committee has met over the last 2 years to classify the channels and the drugs. This report consists of an overview of current thinking, which will be finalized in a document submitted to the IUPHAR (International Union of Pharmacologists) nomenclature committee.


[A case of fatal malabsorption syndrome caused by strongyloidiasis complicated with isosporiasis and human cytomegalovirus infection]. This 54-year-old Korean coal miner suffered from continuous watery diarrhea and weight loss after corticosteroid treatment (beta-methasone, 4 mg daily for 1 week) due to hip-bone fracture in January 1991. Except for the short therapy of steroid, no other histories were contributory. The malabsorption syndrome was aggravated while the case was treated under the impression of amebiasis or intestinal tuberculosis. AIDS antibody test by EIA was negative and quantitative analysis of serum immunoglobulins was in normal ranges. Nine months after the onset of symptoms, the case was diagnosed as malabsorption syndrome caused by complexed and aggravated infection by Strongyloides stercoralis, Isospora and cytomegalovirus in the small intestine, which were proved by stool examination and duodenal biopsy. His clinical course became worse even after high-dosed and prolonged albendazole treatment for strongyloidiasis with supportive fluid therapy. The patient was discharged in hopeless status in November, 1991 and died after one week at home.


Diagnosis of heterozygous familial hypercholesterolemia. DNA analysis complements clinical examination and analysis of serum lipid levels. The concordance of clinical and molecular genetic diagnoses of heterozygous familial hypercholesterolemia (FH) was studied in 65 subjects (10 propositi and 55 first-degree relatives) from 10 families with FH. Nine propositi were carriers of the FH-Helsinki deletion of the low density lipoprotein (LDL) receptor gene, prevalent in the Finnish population, while a new deletion, extending from intron 14 to intron 15 of the LDL receptor gene, was identified in one family. Serum LDL cholesterol levels used in the clinical diagnosis (less than 5.0 mmol/l, not FH; 5.0-5.9 mmol/l, possible FH; greater than or equal to 6.0 mmol/l, FH; limits are 1 mmol/l lower for those less than 18 years) were derived from an authoritative recommendation. Tendon xanthomas constituted an additional criterion. With the DNA analysis as the reference, 55 (85%) subjects could be correctly classified clinically as FH patients or subjects without FH. The remaining 10 subjects were misclassified or were in the "possible FH" category. When the age- and sex-specific 95th percentile LDL cholesterol levels were used instead of the rigid values for both adults and children, the percentage of correct diagnoses rose to 95%. Common genetic polymorphisms of apolipoproteins E and B did not markedly affect LDL cholesterol levels in FH patients, whereas increasing age and obesity were associated with elevated LDL levels. In conclusion, DNA analysis is a valuable adjunct to the diagnosis of FH that is applicable to families with a known mutation of the LDL receptor gene. If DNA methods are not available, age- and sex-specific LDL levels should be used as an aid in the clinical diagnosis of FH.


Tissue reaction to bone-anchored percutaneous implants in rabbits. It has been demonstrated that direct attachment to bony skeletal tissues favors the longevity of percutaneous devices (PDs). This study is focused on improvement of the success rates by using a two-stage implantation technique. For this purpose, cylindrical-shaped PDs were inserted into the tibia of rabbits. The enossal part of the implants was made of Ti6Al4V and was coated with a layer hydroxyapatite (HA). The percutaneous part was made of dense HA. In a first surgical session, the enossal part of the implants was inserted. In a second implantation session, about 3 months later, the percutaneous part was fixed into the enossal part. The implants were left in situ for 5 months after the second session. Clinical and histological investigations were performed. The results confirmed the previous finding--that bone anchoring is effective in the maintenance of a permanent percutaneous passage. On the other hand, it could not be proven that a two-stage implantation procedure improves the success rate of one-stage bone-anchored PDs.


An extremely stable inorganic pyrophosphatase purified from the cytosol of a thermoacidophilic archaebacterium, Sulfolobus acidocaldarius strain 7. A highly active inorganic pyrophosphatase was purified to electrophoretical homogeneity from the cytosol of Sulfolobus acidocaldarius strain 7, an extremely thermoacidophilic archaebacterium. The enzyme has an apparent molecular mass of 80 kDa as estimated by gel permeation chromatography, and showed a 21-kDa polypeptide on SDS-PAGE, suggesting that the archaebacterial enzyme is similar to most of the eubacterial pyrophosphatases rather than eukaryotic ones. The pI = 5.1. The enzyme showed relatively high content of Pro and low content of Ser plus Thr. The optimal pH was 6.5 (at 56 degrees C). From the Arrhenius plot an activation energy of 11.2 kcal/mol was obtained between 37-95 degrees C. The specific activity was 617 mumol Pi release min-1 mg-1 at 56 degrees C. The S. acidocaldarius pyrophosphatase was extremely stable. Complete activity remained after incubation at 100 degrees C for 10 min. No dissociation into subunit or unfolding of polypeptide chain occurred in the presence of 8 M urea. Experiments using guanidine-HCl suggested that the transition between a native tetrameric state and an unfolded state is completely reversible, and essentially independent of any additional factors such as divalent metal cation or dithiothreitol.


Effect of dimethyl sulfoxide on phosphoryl transfer catalyzed by yeast hexokinase. Hexokinase is a phosphotransferase that catalyzes phosphoryl transfer from ATP to glucose much more rapidly than the transfer from ATP to water (i.e., hydrolysis). Dimethyl sulfoxide has opposite effects on these two phosphotransferase activities: it enhances ATP hydrolysis and inhibits glucose phosphorylation. Xylose, a sugar that is non-phosphorylatable by hexokinase, enhances ATPase activity which is additive to activation by dimethyl sulfoxide, indicating that the mechanism of activation by dimethyl sulfoxide is different from that of xylose. These results suggest that it is possible to change the specificity of the enzyme in the presence of dimethyl sulfoxide.


A high-molecular-weight (greater than 8.10(5)) non-collagenous glycoprotein is synthesized by bovine cartilage in vitro. A high-molecular-weight (greater than 8.10(5)) glycoprotein was detected in [3H]glucosamine-labeled bovine cartilage. Extraction with varying amounts of guanidinium chloride showed that the molecule was not tightly bound to other matrix substances. Enzyme digestions identified the molecule as a non-collagenous glycoprotein. This glycoprotein constituted 10-20% of the [3H]glucosamine-labeled macromolecular material that was released into culture medium on the first day after labeling. The 3H-labeled glycoprotein was purified by anion-exchange chromatography, CsCl gradient centrifugation and gel filtration. The purified glycoprotein appeared on an SDS-polyacrylamide gel as one slightly polydisperse band, which could not be reduced by beta-mercaptoethanol.


Heart transplant rejection treated with plasmapheresis. The vascular element of a mixed type of rejection after orthotopic heart transplantation did not respond to steroid therapy in a 30-year-old man. Plasmapheresis was associated with the resolution of the clinical signs of rejection and the microscopic changes of vasculitis in the biopsy specimen. This case suggests that plasmapheresis may be useful in the treatment of acute vascular rejection in heart transplantation.


Activation of phospholipase A2 and beta-thromboglobulin release in human platelets: comparative effects of thrombin and fluoroaluminate stimulation. Several reports have suggested that the activity of platelet phospholipase A2 is modulated by GTP-binding protein(s) whose nature and properties need to be defined. Fluoroaluminate is known to activate G-proteins and this leads to a number of cellular responses including the activation of phospholipases. This paper demonstrates that human platelets, prelabelled with [3H]arachidonic acid, produce free arachidonic acid when stimulated with fluoroaluminate and this effect is time- and dose-dependent. The production of arachidonic acid is not inhibited by neomycin, a PI-cycle inhibitor, but is completely abolished by mepacrine, an inhibitor of both phospholipase A2 and C. At low concentration of fluoroaluminate (10 mM NaF) phospholipase A2 but not phospholipase C is activated. In addition, fluoroaluminate treatment releases beta-thromboglobulin (beta-TG) and this effect is not inhibited by acetylsalicylic acid. Under identical conditions both neomycin and mepacrine suppress the release of arachidonic acid and beta-TG induced by thrombin. Sodium nitroprusside, which increases cGMP levels in platelets, inhibits arachidonic acid liberation and beta-TG release in thrombin-stimulated platelets but has no effect in fluoroaluminate-treated platelets; cGMP was reported to suppress phospholipase C activation. These results are consistent with the hypothesis that, in thrombin-stimulated platelets, the liberation of arachidonic acid and beta-TG are strictly dependent on the activation of phospholipase C. We have also provided evidence for the existence of a phospholipase A2 activated by a G-protein which is independent from the degradation of phosphoinositides and, contrary to phospholipase C, it is not down regulated by cGMP.


Natural anticoagulant pathways in normal and transplanted human hearts. We have studied two natural anticoagulant pathways in normal and in transplanted human hearts. The first is the thrombomodulin pathway. Our immunocytochemical results show thrombomodulin localized to endothelium in heart biopsy specimens before transplantation. This reactivity persists in the absence of cellular rejection, but the infiltration of immune cells is associated with a lack of endothelial thrombomodulin. The second pathway is composed of antithrombin III (ATIII) bound to heparan sulfate proteoglycan (HSPG) molecules on endothelial cells. These ATIII-HSPG complexes bind and inactivate thrombin at the endothelial surface. Our immunocytochemical results show ATIII localized to endothelium in heart biopsy specimens before transplantation. This reactivity is present in the absence of vascular rejection as defined by either angiography or microscopy. The absence of thrombomodulin and ATIII is always associated with fibrin deposition within the microcirculation. Thrombomodulin and ATIII pathways appear to be independent, for cellular rejection often is associated with thrombomodulin-negative ATIII-positive endothelium, and vascular rejection often is associated with thrombomodulin-positive ATIII-negative endothelium. Cytokines from activated macrophages down-regulate endothelial thrombomodulin without generally affecting the ATIII-HSPG pathway. Immunosuppressive therapy depletes cytokine-producing cells that affect thrombomodulin, but there presently is no therapy to protect endothelium in vascular rejection. It is possible that heparin could interact with endothelium and bind ATIII to maintain a state of thromboresistance.


Anterior spinal artery syndrome of the cervical hemicord. Three patients developed signs of a unilateral cervical cord lesion 6 to 36 h after the acute onset of severe cervico-brachial pain. The neurological deficit progressed over 6 to 18 h. On the painful side a central Horner's syndrome, a hemiparesis with plegia of the hand, and a slight pallhypaesthesia were found. On the opposite side thermhypaesthesia and hypalgesia were noted with a level at the dermatome C5 or C6. T2-weighted MR images revealed in one patient a small area of increased signal intensity restricted to one half of the cervical cord, and electromyography in another patient showed after 6 months evidence of segmental chronic denervation. Both abnormalities were found at the clinically expected level. The findings are consistent with a small infarction of the cervical cord in the perfusion territory of a central (sulco-commissural) artery, a duplicated anterior spinal artery or an anterior spinal branch of the vertebral artery.


Protein kinases and the androgen-induced proliferation of accessory sex organ smooth muscle. The possible role of second messenger systems in androgen-dependent smooth muscle proliferation was investigated. Focusing on the hormone-sensitive guinea pig seminal vesicle, we analyzed changes in protein kinase C (PKC) and cAMP-dependent type I and II protein kinases during the androgen-dependent smooth muscle proliferation of puberty, as well as in the transition to the nonproliferative state of the adult. The androgenic sensitivity of the cAMP-dependent type I and II protein kinases and the cAMP-dependent phosphorylations of soluble muscle proteins did not correlate with the qualitative change in the androgenic sensitivity of the prepubertal vs. adult animals. In contrast to the cAMP-dependent protein kinases, regulation of the soluble and particulate forms of PKC corresponded to the androgen-induced smooth muscle proliferation. That is, in the seminal vesicle muscle of prepubertal castrated animals, androgen treatment reduced both the soluble and particulate forms of PKC during the increase in smooth muscle DNA synthesis, and in adult seminal vesicle smooth muscle, which was resistant to androgen-induced proliferation, both forms of the enzyme were resistant to androgenic stimulation. It is concluded that PKC may be a component of an autocrine mitogenic mechanism involved in the coupling and uncoupling of androgen-induced smooth muscle proliferation.


Bovine retained placenta: effects of collagenase and hyaluronidase on detachment of placenta. A significant percentage of cows (11%) fail to release the placenta within 12 h postpartum. Failure of collagen breakdown seems to be related to the retention of placentas. Sections of placentomes incubated with bacterial collagenase caused an increase in placentome proteolysis (6.6-fold) and placentome collagenolysis (94-fold) within 4 h in a dose-related fashion (r = 0.94). Injections of collagenase (825 U/cc) into the placentomes, via umbilical vessels, decreased the cotyledon-caruncle binding force (determined by manometry) to 30 +/- 5 mm Hg from 97 +/- 2 mm Hg, and increased proteolysis by 42% within 8 h (r = -0.95). Hyaluronidase at various concentrations (400-8 250 U/cc) and at various incubation times (up to 8 h) was not effective. Hyaluronidase (825 U/cc) and collagenase (825 U/cc) were not synergistic in loosening cotyledon-caruncle attachment. A single 15-min collagenase pulse, given prior to perfusion with collagenase-free blood, was as effective in loosening cotyledon attachment as was a sustained 2-h perfusion of blood with collagenase added. It was concluded that collagenase caused collagenolysis and loosening of cotyledon from caruncle, but collagenolysis and cotyledon-caruncle separation were not facilitated by the presence of hyaluronidase.


Multifactorial regulation of prostaglandin synthesis in preovulatory goldfish ovarian follicles. Goldfish preovulatory ovarian follicles (prior to germinal vesicle breakdown) were utilized for studies investigating the actions of activators of different signal transduction pathways on prostaglandin (PG) production. The protein kinase C (PKC) activators phorbol 12-myristate 13-acetate (PMA; 100-400 nM), 1-oleoyl-2-acetylglycerol (5 and 25 micrograms/ml), and 1,2-dioctanoylglycerol (10 and 50 micrograms/ml) stimulated PGE production; the inactive phorbol 4 alpha-phorbol didecanoate, which does not activate PKC, had no effect. Calcium ionophore A23187 (0.25-4.0 microM) stimulated PGE production and acted in a synergistic manner with activators of PKC. Although produced in lower amounts than PGE, PGF was stimulated by PMA and A23187. The direct activator of phospholipase A2, melittin (0.1-1.0 microM), stimulated a dose-related increase in PGE production, whereas chloroquine (100 microM), a putative inhibitor of phospholipase A2, blocked basal and PMA + A23187-stimulated PGE production. Several drugs known to elevate intracellular levels of cAMP including the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (0.1-1.0 mM), forskolin (10 microM), and dibutyryl cAMP (dbcAMP; 5 mM) attenuate PMA + A23187-stimulated PGE production. Melittin-stimulated production of PGE was inhibited by dbcAMP, suggesting that the action of cAMP was distal to the activation of phospholipase A2. In summary, these studies demonstrate that activation of PKC and elevation of intracellular calcium levels stimulate PG production, in part, through activation of phospholipase A2. The adenylate cyclase/cAMP signalling pathway is inhibitory to PG production by goldfish ovarian follicles.


Biphasic effect of calcium on luteinizing hormone-stimulated cyclic adenosine 3',5'-monophosphate production in granulosa cells of the fowl (Gallus domesticus). Both cAMP and Ca2+ play important roles in the steroidogenic action of LH in hen granulosa cells. However, the interaction of these intracellular messengers is not fully understood. In the present study we used two calcium ionophores (ionomycin and A23187), as well as trifluoperazine (TFP), an inhibitor of calmodulin, to investigate LH- and forskolin-induced cAMP production in granulosa cells isolated from the largest (F1) preovulatory follicle of White Leghorn laying hens. Between 0.1 and 1.0 microM, both ionophores significantly potentiated cAMP responses to LH in the presence of 0.1 mM extracellular Ca2+. When calcium was omitted from the medium, ionophores had no effect. When either calcium was raised above 1 mM, or the concentration of ionophores was increased above 1 microM, LH-induced cAMP production was drastically inhibited. In the presence of 0.5-2.0 mM calcium, A23187 inhibited forskolin-promoted cAMP synthesis. TFP, while having no effect on basal cAMP, suppressed LH-induced responses and the potentiating effect of ionomycin. It is concluded that for full activation of the adenylate cyclase/cAMP system by LH, Ca-calmodulin is required at a site upstream from the catalytic component of the enzyme. However, high intracellular Ca2+ and/or other effects of ionophores (such as uncoupling of oxidative phosphorylation) inhibit LH-induced cAMP production.


Phagocytosis of immunoglobulin G and C3-bound human sperm by human polymorphonuclear leukocytes is not associated with the release of oxidative radicals. Antisperm antibody (ASA)- and complement (C)-mediated immune injury to human sperm is thought to be caused in part by phagocytic neutrophils. To investigate this process, we co-cultured purified human polymorphonuclear leukocytes (PMN) with swim-up sperm in the presence of ASA-positive and ASA-negative sera and assayed for PMN respiratory burst activity, monitored by the release of superoxide anion (O2-) and hydrogen peroxide (H2O2). Phorbol myristate acetate (PMA) and opsonized zymosan were used as positive controls. Phagocytosis of ASA-positive and C-bound sperm by PMN did not enhance O2- production when compared to incubation of sperm with ASA-negative sera. Phagocytosis of ASA-positive and C-bound sperm also resulted in minimal release of H2O2 when compared with ASA-positive and C-negative sperm that were not phagocytosed. In contrast, PMN were maximally stimulated to release O2- in response to either opsonized zymosan or PMA. The kinetics of PMA-induced O2- release was unaffected by the presence of ASA-positive and C-bound sperm. Cytocentrifuge preparations of PMN incubated with ASA-positive and C-bound sperm revealed limited O2- release at the site of PMN/sperm contact. These results indicated that 1) phagocytosis of motile sperm by PMN requires the binding of both ASA and C to the sperm surface; 2) phagocytosis of ASA-positive and C-positive sperm by PMN fails to release reactive oxygen species; and 3) metabolic processes associated with PMN respiratory burst activity may not be coupled to the ingestion of ASA-positive and C-bound sperm.


Treatment of interstitial pneumonitis due to cytomegalovirus with ganciclovir and intravenous immune globulin: experience of European Bone Marrow Transplant Group. Data on 49 allogeneic bone marrow transplant (BMT) recipients who developed interstitial pneumonia due to cytomegalovirus (CMV) were collected retrospectively. All patients were treated with ganciclovir and high doses of intravenous immune globulin, although types of immune globulins and schedules of treatment varied. Seventeen (35%) of 49 patients responded to treatment. Thirty days after the diagnosis of interstitial pneumonia, the survival rate among patients was 31%. CMV was detected in 81% of patients on whom autopsies were performed. The survival rate among patients who received total body irradiation (TBI) was significantly lower (11 [27%] of 41) than that among patients who did not receive TBI (six [75%] of eight; odds ratio = 12.3; P = .009). No other factor, including age, grade of graft-versus-host disease, types and dose of immune globulin used, or dose of ganciclovir, was correlated to survival. These results show that although survival of allogeneic BMT recipients with CMV interstitial pneumonia has improved, more than one-half of the patients still died of pneumonia. Thus, both prophylaxis for and treatment of CMV infection must be improved.


Penicillium marneffei infection in patients infected with human immunodeficiency virus. From June 1990 to August 1991, 21 patients infected with the human immunodeficiency virus (HIV) presented with systemic mycosis caused by Penicillium marneffei. Between August 1987 and August 1991, only five patients were observed who had P. marneffei infection but not HIV infection. The clinical presentation included fever, cough, and generalized papular skin lesions. For 11 of these 21 patients, the presumptive diagnosis of P. marneffei infection could be made by microscopic examination of Wright's-stained bone marrow aspirate and/or touch smears of skin specimens obtained by biopsy several days before the results of culture were available. Initial clinical response to treatment with either parenteral amphotericin B or oral itraconazole was favorable in most patients. Epidemiological and clinical evidence suggest that this systemic mycosis is caused by an important opportunistic pathogen and that it should be included in the differential diagnosis of AIDS, at least for countries in areas of endemicity, i.e., Southeast Asia and China.


Herpesviruses--immune escape artists?

Herpesviruses--immune escape artists? Viral persistence depends on the successful avoidance of the host's immunologic surveillance system. This review, which focuses specifically on the herpesviruses, delineates several possible strategies for evading or delaying the immune response. One strategy common to all herpesviruses is the establishment of latency, a state in which the virus may be partially or even completely hidden from the immune system. Other proposed mechanisms of immune evasion include interaction of the virus with components of the humoral immune system, virus-induced modulation of cell-surface recognition structures, and virally mediated interference in antigen processing. Additional strategies include molecular mimicry and the ability of one particular herpesvirus to encode an immunosuppressive cytokine. Although a detailed understanding of the molecular mechanisms of herpesvirus-mediated immune evasion is currently lacking, future studies should identify those critical interactions between host and virus that may prove amenable to therapeutic intervention.


Insulin resistance and endogenous digoxin-like factor in obese hypertensive patients with glucose intolerance. Hypertensive obese subjects with glucose intolerance have hyperinsulinaemia, insulin resistance and intracellular cation imbalance resulting in increased sodium content. The aim of our study was to assess in these patients plasma levels of endogenous digoxin-like factor (EDLF), an inhibitor of the sodium-pump mechanism. We studied 14 hypertensive and 12 normotensive subjects with obesity and glucose intolerance for fasting blood glucose, and plasma insulin, C-peptide and EDLF levels: the two groups were matched for age and BMI and were studied after a 2-week wash-out period from hypotensive drugs. Compared with normotensives, hypertensive subjects had higher plasma insulin levels, a greater immunoreactive insulin/C-peptide ratio, a lower glucose/insulin ratio and higher plasma EDLF levels. Our results confirm that among obese people with glucose intolerance, hypertensives are more hyperinsulinaemic and insulin-resistant than normotensives and indicate that the intracellular cation imbalance in these patients may be attributable, at least in part, to EDLF.


Hepatitis B vaccination: an important method of preventing HBV-related hepatocellular carcinoma. Hepatocellular carcinoma (HCC) is a multifactorial disease which requires multiple strategies for its prevention, including immunization against hepatitis type B. Given the link between chronic HBV infection and HCC, vaccination against hepatitis B preventing an initial infection and the development of chronic carrier state is also likely to prevent HCC. The percentage of HCC cases attributable to HBV, which depends on both relative risk for HCC associated with the HBsAg carrier state and prevalence of HBsAg carriers in a particular population, is much higher in HBV hyperendemic countries than in those at low endemicity. Thus prevention of HCC by hepatitis B vaccination should have the highest benefit in areas at high level of endemicity. Unfortunately, the poor health-care resources make the establishment of mass-immunization campaigns against HBV difficult in most of these countries.


Embolization and chemoembolization for hepatocellular carcinoma. A check on the rationale for embolization and chemoembolization for hepatocellular carcinoma and a revisit to the background of the anatomy of blood supply to the liver is discussed. The technique of the embolization is different and mostly in the range of home made configuration. There is no data concerning the stability of different choices such as the popular mixture of cytotoxic agents and lipiodol. Therefore the authors have preferred the use of microencapsulated Mitomycin believing that this special formulation of the drug can attain the present best reproducibility. The survival of 32 treated patients was longer than the control group. Twelve, 36 and 60 month survival was 70%, 45% and 15% vs 37%, 0%, and 0% respectively.


Liver resection for hepatocellular carcinoma in cirrhotic patients. The results of a multicentric retrospective analysis of 265 liver resections for hepatocellular carcinoma in cirrhotic patients have been collected and assessed. Overall operative mortality was 9.1%. The actuarial 3-year survival was 46.5%. Better results were obtained in patients with encapsulated lesions and alphafetoprotein levels less than 400 ng/dl. Tumoral recurrence, which was the main cause of late death, was related to alphafetoprotein higher levels and to the type of surgery performed: tumor resection with short tumour free margin had the highest recurrence rate. Results were significantly better in those centers where intraoperative ultrasonography was systematically employed.


Percutaneous ethanol injection of hepatocellular carcinoma: survival after 3 years in 70 patients. Percutaneous ethanol injection (PEI) under ultrasound control is a new therapeutic procedure for cirrhotic patients with small hepatocellular carcinoma (HCC). Our experience included 70 patients with 110 lesions (range 0.8-5 cm). Follow-up ranged from 5 to 70 months (mean: 25 months). No serious complications occurred after 930 treatments. The 1, 2, 3-year survival rates (Kaplan-Meier method) were: 96, 84, 72% for single lesions less than 5 cm and 94, 84, 25% for multiple lesions. In our opinion and in consideration of the results of other authors obtained with PEI and with surgery, single lesions less than 5 cm found in Child's A and B patients with high surgical risk or with some adverse prognostic factors for surgery, could be an indication for PEI.


Computed tomography and magnetic resonance imaging in diagnosing hepatocellular carcinoma. The evaluation of hepatocellular carcinoma (HCC) is based upon ultrasonography (US) which has proved to have a high sensitivity and is also extremely useful in guiding the percutaneous needle biopsy. The main role of computed tomography (CT) and magnetic resonance imaging (MRI) is to supplement US in evaluating the extent of HCC. The Authors discuss the different techniques of examinations of the liver both for CT and MRI as far as the modalities of contrast enhancement, site of injection, and type of contrast agents are concerned. The differences between low field and high field magnets are also discussed. The main CT and MRI findings are illustrated, depending upon the technique of examination. Finally the role of these techniques is discussed. Based upon personal experience and the data in CT literature, and if performed with updated technology and intraarterial injection (lipiodol), CT is the method of choice in order to supplement US in the evaluation of HCC.


Arteriography in diagnosing hepatocellular carcinoma. The diagnostic role of arteriography in the evaluation of hepatocellular carcinoma is pointed out. In the identification of large lesions angiography achieves a sensitivity of 100%. However, when evaluating small lesions (under 5 or 3 cm in diameter), the sensitivity is reported to be 82-89%. To increase the sensitivity, different special techniques have been developed which require angiographic placement of a catheter. These include CT arteriography, CT arterial portography and CT after intraarterial injection of iodized oil. Particularly with the two latter techniques, the sensitivity in identification of small hepatocellular carcinoma reaches a value of 91-95%. Angiography can also be proposed for the characterization of neoplastic lesions in cirrhotic livers. Its specificity is 72% in the differential diagnosis of hepatocellular carcinoma and adenomatous hyperplastic nodules. Small grade I hepatocellular carcinoma can simulate this benign lesion at angiography, as well as at histology. Finally, angiography may still play a role in the preoperative staging of hepatocellular carcinoma both for the diagnosis of satellite lesions and for the identification of vascular invasion.


The natural history of hepatocellular carcinoma. Prospective studies of patients with cirrhosis who are at risk for developing hepatocellular carcinoma (HCC), have greatly improved our understanding of the natural history of this tumour. Despite many studies, the controversy of whether HCC originates from one or more than one clone of transformed liver cells is still unsettled. The natural history of HCC in many instances is an extension of that of the underlying cirrhosis, although the course of the tumour disease may vary from patient to patient and in different geographical areas. In Oriental and southern European patients with HCC, a longer time may elapse between diagnosis and death, than in African blacks. The information about tumour development in non-cirrhotic liver is scanty, due to the little known risk factors for this type of tumour which has impeded performing prospective cohort studies of high risk patients.


Itraconazole in antifungal therapy.

Itraconazole in antifungal therapy. This overview compares and contrasts the pharmacotherapy of itraconazole with that of other antifungal agents. Primary literature on itraconazole was identified through a medical literature search from 1976 through 1991. This search included journal articles, abstracts, conference proceedings, and reports of animal and human research published in the English language. All primary literature was reviewed regardless of the study design or outcome. Literature evaluations of efficacy were ranked using a literature rating scale (Dalen JE, Hirsh J. Arch Intern Med 1986;146:462-72), which was slightly modified to include case reports and observations. All data were collected and represented with a primary focus on itraconazole's mechanism of action, pharmacokinetics, clinical efficacy in systemic mycotic infections, drug interactions, and adverse reactions. All articles were referenced in the final data presentation unless grouped data had been accurately reviewed and published. Despite the paucity of controlled comparative trials with itraconazole in patients with deep mycoses, results on efficacy are encouraging. It is still unclear what role itraconazole will have in the prophylaxis of fungal infections in immunocompromised hosts. The favorable pharmacokinetic profile permits once- or twice-daily administration and itraconazole appears to be safe and well tolerated. Itraconazole should prove to be a useful replacement for ketoconazole on hospital formularies. This recommendation is based on itraconazole's greater apparent safety and efficacy. Reevaluation of this agent will be necessary upon the release of newer imidazoles and triazoles.


Radioiodination of S-type lipopolysaccharide. Possibilities and limitations of labelling of the carbohydrate chain by periodate oxidation. S-type Lipopolysaccharide (LPS) from E. coli O 139:K 82 (B) was radiolabelled by coupling 125I-tyramine to the carbohydrate chain of LPS after oxidation of sugar residues carrying vicinal hydroxy groups, with sodium metaperiodate. The specific activity amounted to 1.3 kBq/microgram LPS. As LPS is a mixture of molecules with different carbohydrate chain lengths that are associated to large micelles of high kinetic stability, the preparation and the properties of the tracer exhibit some peculiarities. Most important, partial cleavage of the carbohydrate chains of LPS is caused on radioiodination by the procedure described. As a consequence, the tracer differs markedly from the starting LPS with respect to carbohydrate chain length distribution. Another feature of special interest is the variation of the specific activity (radioactivity/unit mass) for LPS molecules of different carbohydrate chain lengths. Because of the kinetic stability of LPS micelles, the equilibration of radioiodinated and non-radioactive LPS aggregates requires their solubilization by a detergent and its subsequent removal by dialysis. It could be demonstrated that short-chain LPS molecules predominantly associate to larger micelles than long-chain molecules. Regardless of these restrictions, the 125I-LPS proved to be useful for certain analytical purposes.


Effects of white light on the pineal gland of the chick embryo. Chick embryos were directly exposed to a source of white light during incubation and sacrificed before hatching. The light caused a number of teratological effects such as high mortality, delay in development, celosomy, hepatomegaly, auricular dilation and micrognatia. The pineal gland of the illuminated embryos showed an increase in number and size of the intracytoplasmic lipid droplets of the follicular pinealocytes. These findings suggest that the pineal gland of the chick embryo is sensitive to light.


Morphodynamic response of the rat light pinealocytes to an injection act. Implication of beta-adrenoreceptors. The implication of beta-adrenoreceptor-mediated noradrenalin action on the reactive morphodynamic response of light pinealocytes (LP) to an injection act, conceived as a short-lasting stress attack, is reported. An injection act, realized by saline-injection, was manifested in the occurrence of clustered "dusk" and "bright" cells--the representative forms of functionally activated LP. The time-related incidence of these cells entities--the appearance of "dusk" and "bright" cells at 5 min, transitory domination of "bright" cells and the nadir of "dusk" cells at 20 min, sporadic recognition of "bright" cells, lack of "dusk" cells at 45 min and the absence of both cell forms at 180 min--displayed that LP-reactive response promptly appeared and rapidly ceased. The injection of beta-adrenoreceptor antagonist, propranolol, did not considerably alter the pattern of LP-reactive response proper for saline-injected rats at 5 min. The constant presence of "dusk" and "bright" cells, structurally changed in all the following time periods under investigation, showed that this drug disordered the course of LP-reactive response to an injection act. On the contrary, LP of resting state was not found to be affected. Estimating functionally, the present results indicate that beta-adrenoreceptor-mediated noradrenalin action is required to promote, maintain and accomplish the LP-reactive response to a short-lasting stress inducement.


ACTH-induced ultrastructural changes in the zona fasciculata of the hamster adrenal cortex. Are intraadrenal thrombi regulators of corticosteroid secretion? Correlated stereological and functional studies were performed on the effect of massive ACTH doses on adrenal cortex of the female hamster. ACTH resulted in a marked increase in adrenal gland weight at day 6 of treatment followed by a drop at day 9. Stereology showed significant enlargement of the zona fasciculata (ZF) cells with the highest value at day 6 and subsequent drop at day 9 of treatment. This hypertrophy was due to a notable increase in the volume of mitochondrial, SER, Golgi apparatus and lipid droplet compartments. Cortisol secretion by adrenal slices and homogenates was also highest at day 6 of ACTH administration and notably lower at day 9. At day 6 of ACTH treatment in outer ZF thrombi were seen. In their vicinity the subendothelial space was dilated and endothelial cells dissociated from the basal lamina. Numerous erythrocytes were also visible among dissociated ZF cells. At day 9 of experiment in outer part of ZF numerous spaces devoid of parenchymal cells appeared. The earlier authors considered the "empty spaces" or "holes" in hyperstimulated adrenal cortex as a sign of holocrine secretion of steroid hormones. The present findings enable us to introduce a new hypothesis on the development of these spaces. In our opinion in hyperstimulated adrenal cortex numerous thrombi may be formed leading thus to the degeneration of adrenocortical cells. Thus, the appearance of the "empty spaces" or "holes" in the gland is not connected with the holocrine secretion but with the regulation of the number of secretory cells in adrenal cortex by the thrombi-dependent mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)


Axonal and transsynaptic (transneuronal) spread of Herpesvirus simiae (B virus) in experimentally infected mice. In order to study the pathogenesis of B virus infection of the nervous system, newborn and young mice were inoculated by four different routes: 1. Intramuscular (i.m.) in the forelimb; 2. I.m. in the hindlimb; 3. Subcutaneous (s.c.) in the abdominal wall; 4. Intraperitoneal (i.p.). Spread of virus was followed by immunohistochemical demonstration of viral antigen in tissue sections of the peripheral and central nervous system. Three distinct patterns emerged: 1. After i.m. limb inoculations, virus progressed along the ipsilateral dorsal column, the bilateral spinothalamic and bilateral spinoreticular systems and along central autonomic pathways. 2. After s.c. inoculation, the dorsal column was spared, otherwise the spread was similar to that following i.m. inoculations. 3. After i.p. inoculation, virus spread in the spinal cord bilaterally, mainly along spinothalamic and central autonomic pathways. The peripheral motoneurons were conspicuously spared, even in the i.m. inoculation mode. In the brain stem, B virus antigen appeared bilaterally, at multiple sites. In the cerebrum, virus infected cells appeared first in the thalamus, hypothalamus and the motor cortex. The mode of spread from spinal levels was mainly orthograde along the ascending systems (dorsal columns, spinothalamic, spinoreticular tracts), but also retrograde along descending systems (pyramidal tract, central autonomic pathways). Oligosynaptic systems transmitted virus more quickly than the polysynaptic ones. In the involvement of various neuronal systems in virus spread, a certain selectivity, sparing the peripheral motoneuron and the cerebellar systems, could be assessed.


PTH stimulates the proliferation of TE-85 human osteosarcoma cells by a mechanism not involving either increased cAMP or increased secretion of IGF-I, IGF-II or TGF beta. Injections of parathyroid hormone (PTH) result in increased bone formation in several species. Work in our laboratory and others has shown a stimulation of bone cell proliferation and growth factor production by PTH. Our purpose was to study the effects of PTH on a human bone cell line using TE-85 human osteosarcoma cells as a model. After 24 h treatment, PTH caused an increase in cell proliferation as measured by cell counts and [3H]-thymidine incorporation. Proliferation was not inhibited by an anti-transforming growth factor beta (TGF beta) antibody which could abolish stimulation by exogenous TGF beta. PTH did not stimulate cAMP production, alkaline phosphatase activity or production of insulin-like growth factors I or II (IGF-I or IGF-II) in TE-85 cells. Although basal TE-85 proliferation was slowed by incubation with the calcium channel blocking agent verapamil, PTH still caused an increase in growth rate. We conclude that PTH directly stimulates TE-85 proliferation via a mechanism not involving increased adenylate cyclase activity or increased secretion of IGF-I, IGF-II or TGF beta and may stimulate bone formation in vivo by activating some other mitogenic signal to increase bone cell proliferation.


Glomus tumour: an analysis of 43 patients and review of the literature. Glomus tumours are hamartomas derived from the normal glomus body. A total of 43 glomus tumours have been seen over the past 18 years, and are reviewed. Their clinical presentations and pathology are analysed and the continuing difficulty in diagnosis is examined.


Potentiation of nicotinic transmission in the rat superior cervical sympathetic ganglion: effects of cyclic GMP and nitric oxide generators. The efficacy of nicotinic transmission in the rat superior cervical ganglion in vitro (24-26 degrees C) was estimated by extracellular recording of the postganglionic compound action potential response to stimulation of the preganglionic nerve at a slow rate (one shock every 60 s). Atropine (2 microM) was included to block muscarinic transmission, and hexamethonium (200-250 microM) was used to produce a submaximal response sensitive to potentiation and inhibition of nicotinic transmission. Upon exposure to 1-100 microM 8-bromo-guanosine 3',5'-cyclic monophosphate (8-Br-cGMP), nicotinic transmission was potentiated by 6 +/- 1% (n = 4) to 89 +/- 5% (n = 5) in a dose-dependent manner. 8-Bromo-adenosine 3',5'-cyclic monophosphate (8-Br-cAMP, 10-100 microM) also potentiated nicotinic transmission (3.8 +/- 0.3% (n = 3) to 43 +/- 4% (n = 3)). However, 8-Br-cGMP was at least 2-fold more effective than 8-Br-cAMP. Sodium nitroprusside (0.1 microM to 1 mM) and sodium azide (0.1-100 microM) were used to stimulate the formation of endogenous cGMP52. Nicotinic transmission was potentiated by these substances also. The response was increased by 3.4 +/- 0.7% (n = 4) to 32 +/- 2% (n = 5) upon exposure to 0.1-100 microM sodium nitroprusside, and by 5.5 +/- 0.9% (n = 3) to 18 +/- 4% (n = 4) upon exposure to 0.1-100 microM sodium azide. Ferricyanide ion (10-100 microM) appeared to be ineffective, as would be expected if the effect of nitroprusside was due to the nitric oxide rather than the cyanide or ferric moieties.(ABSTRACT TRUNCATED AT 250 WORDS)


Development of inositol 1,4,5-trisphosphate 3-kinase immunoreactivity in cerebellar Purkinje cells in vivo and in vitro. Development profiles in vivo and in vitro of inositol 1,4,5-trisphosphate 3-kinase (IP3K) were investigated immunohistochemically in the cerebellar Purkinje cells. In in vivo preparations of rat cerebellum, IP3K immunoreactivity appeared in Purkinje cell bodies and dendrites shortly after birth, increased rapidly by postnatal day 5, and was subsequently confined to their dendritic processes by day 20. The appearance and shift of IP3K immunoreactivity in Purkinje cells showed an identical time course even when Purkinje cells were placed under culture conditions commencing on day 0, suggesting that Purkinje cells have their own biological clock on the expression of IP3K in the absence of external influences.


Escape from inhibition of synaptic transmission during in vitro hypoxia and hypoglycemia in the hippocampus. Electrophysiological recordings were made from rat hippocampal slices exposed to in vitro ischemic conditions in which the superfused medium is hypoxic and lacking glucose. Under these conditions, the evoked population spike recorded in CA1 is initially depressed and then transiently returns prior to an anoxic depolarization. This transient return in synaptic function under ischemic-like conditions also occurs if the population spike is inhibited by pretreatment with adenosinergic agonists or with the gamma-aminobutyric acid (GABA)B agonist, baclofen.


Suppression of presynaptic calcium currents by hypoxia in hippocampal tissue slices. We tested the hypothesis that suppression of inward calcium current in presynaptic terminals is the cause of failure of synaptic transmission early during cerebral hypoxia. Postsynaptic responses in CA1 zone of hippocampal tissue slices were blocked either by the combined administration of 6,7-dinitroquinoxaline-2,3-dione (DNQX) and 3-((+-)-2-carboxypiperazine-4-yl)-propyl-1-phosphonic acid (CPP) or by lowering extracellular calcium concentration ([Ca2+]o). Repetitive orthodromic activation of central neurons caused transient decrease of [Ca2+]o (measured by ion selective microelectrodes) in neuropil, attributable to influx of Ca2+ in presynaptic terminals. Presynaptic [Ca2+]o responses were rapidly and reversibly suppressed when oxygen was withdrawn from hippocampal tissue slices. The 'resting' baseline level of [Ca2+]o declined at first gradually, then precipitously as in spreading depression (SD). Presynaptic volleys during high frequency train stimulation were also depressed somewhat before SD began. We conclude that (1) presynaptic Ca2+ currents fail during hypoxia, perhaps because 'resting' intracellular free Ca2+ activity is increased and, in part, also because of partial failure of presynaptic impulse conduction; (2) the influx of Ca2+ into brain cells in hypoxic spreading depression is not mediated by glutamate/aspartate dependent channels.


[Polymerase chain reaction in the detection of hepatitis B virus carriers with sole positive anti-hepatitis B core antigen]. To assess the prevalence of serum HBV DNA in the individuals with positive anti-HBc alone, seroepidemiologic investigations in three populations were carried out with polymerase chain reaction. The detection of sole anti-HBc were 21 of 294 (7.1%), 193 of 1995 (9.7%) and 23 of 377 (6.1%) individuals respectively. Among the anti-HBc positive sera, HBV DNA was detected in 6 (28.6%), 68 (35.2%) and 7 (30.4%) respectively. Follow-up observations showed that 83.8%, 93.1% and 71.4% still remained HBV DNA positive respectively. These might be reasonably regarded as chronic asymptomatic HBV carriers with undetectable HBsAg.


[Quantitative direct gas-liquid chromatography for rapid diagnosis of anaerobic infections]. Quantitative direct gas-liquid chromatography (GLC) was performed on 74 specimens of surgical infections and relevant findings were compared with cultural results. In the present study, significant amounts of multiple volatile fatty acids (VFA) and/or succinate were found as markers of anaerobic infections. At least one of the VFA (propionic, isobutyric, butyric, isovaleric or valeric acid) greater than or equal to 0.1 mumol/ml and/or succinate greater than or equal to 0.3 mumol/ml was strong evidence for anaerobic infections. More than 3.5 mumol/ml of butyric acid was interpreted as indication of the presence of Fusobacterium spp. Succinate greater than or equal to 0.3 mumol/ml was associated with the presence of Bacteroides fragilis.


[Preliminary study on the relation between serum growth hormone and 1,25-dihydroxyvitamin D3 in acromegalic patients]. Eight acromegalic patients were treated with SMS 201-995 for 12 months. At the 3rd, 6th and 12th month from the beginning of treatment, blood samples of these patients were taken for determination of growth hormone (GH), 1,25-(OH)2D3, SMC, calcium and phosphorus. The serum levels of GH decreased significantly at the 3rd, 6th and 12th months compared with the basal levels before treatment (P less than 0.01, P less than 0.05 and P less than 0.05 respectively). The concentrations of serum 1,25-(OH)2D3 and SMC also decreased significantly. Mean serum GH, 1,25-(OH)2D3 and SMC levels were reduced from 34.8 +/- 10.7 to 15.5 +/- 5.9 ng/ml (P less than 0.05), 76.2 +/- 2.7 to 45.9 +/- 8.3 pg/ml (P less than 0.05) and 8.5 +/- 1.2 to 4.4 +/- 1.0 U (P less than 0.05) respectively at the 12th month after treatment. Serum phosphorus levels reduced from 4.7 +/- 0.4 to 4.1 +/- 0.2 mg% (P less than 0.05) at the 6th month after treatment, but there was no change in the serum levels of total calcium. The treatment with SMS 201-995 directly evoked a reduction of serum GH in acromegalic patients and at the same time, the levels of serum SMC and 1,25-(OH)2D3 were decreased. The results suggest that GH may stimulate indirectly the production of 1,25-(OH)2D3 through the mediation of SMC.


[Visual and computer evaluation of diazepam-induced EEG changes in epilepsy for localization of epileptogenic lesion]. Diazepam-induced EEG changes were analyzed both visually and by computer in epilepsy for localization of epileptogenic lesion. The results demonstrated that changes in beta spectral were diminished in 11/24 or 15/25 cases, but regionally increased in 3/24 or 3/25 patients overlying the epileptogenic lesion or area of EEG abnormalities. Diazepam-induced delta spectral power was usually increased (9/24), but occasionally focally reduced (3/24) over the lesion. The percentage of positive findings for localization was 38% with visual analysis and 46% with spectral analysis in beta spectral power, 38% in delta spectral power, and 67% with the analysis in combined beta with delta, and 83% with four independent tests. Spectral analysis was more sensitive than visual analysis, and it was a useful supplement to visual analysis, to general EEG evaluation, and to CT. Finally, the study emphasized that a localizing diagnosis to define the epileptogenic lesion should be based on the combination of a multitude of independent tests.


[Effect of cordyceps sinensis on cellular immunity in rats with chronic renal insufficiency]. Animal model of chronic renal failure (CRF) was induced in wistar rats by 5/6 nephrectomy. Half of the rats were treated with Cordyceps sinensis (CS) in form of decoction. It was found that CS has mitogenic effect on spleen lymphocytes, and is capable of increasing the production of IL-2 from splenocytes of the CRF rats. IL-2 absorbency of the splenocytes was promoted by CS. CS also exhibited such therapeutic effects on CRF animal as to decrease the level of BUN and serum creatinine and to increase the level of hemoglobin. These results indicate that CS has a regulative effect on cellular immunity in CRF rats.


[Effects of female sex hormones on atrial natriuretic factor gene expression in rats]. To clarify the effects of female sex hormones on ANF gene expression, emasculated and intact female rats were subcutaneously injected with estradiol (E2), progesterone (P) and olive oil solvent (O) respectively, once a day for 7 days. The relative rANF-mRNA contents of rat atria were measured by molecular hybridization, and rANF-cDNA was labeled with alpha-32P as probe. The groups and hormone doses per 100 g body weight were: (1) sham, 0.2 ml O; (2) rats were bilaterally ovariectomized(OVX), 0.2 ml O; (3) OVX, 80 ng E2; (4) OVX, 80 micrograms E2; (5) OVX, 0.2 mg P; (6) OVX, 2 mg P; (7) OVX, 80 ng E2, 0.2 mg P. The results showed that the rANF-mRNA of group (2) was the lowest, it was 75% of that in group (1), while the rANF-mRNA levels in groups (3)-(7) were 1.1, 2.0, 1.9, 2.3 and 2.1 folds of that in group (2) respectively. The results revealed that E2 and P increased ANF gene expression. Their effects were associated with the dosage. The two may have synergistic actions. Physiological amount of E2 and P may maintain suitable level of rANF-mRNA. This experiment suggested that female sex hormone may have dual effects in body fluid balance.


[Determination of serum creatinine kinase MM isoforms in assessing reperfusion after acute myocardial infarction]. Creatine kinase(CK), aspartate aminotransferase (AST), alpha-hydroxybutyrate dehydrogenase (HBDH), lactate dehydrogenase (LD) and LD isoenzymes, CK-MB isoenzymes and CK-MM isoforms were measured in 17 acute myocardial infarction (AMI) patients treated with thrombolysis resulting in reperfusion and 2 not resulting in reperfusion as well as 71 treated conventionally to assess reperfusion. The results showed that the peak of the ratio of MM3 to MM1 was attained significantly earlier in patients with reperfusion than in those conventionally treated and those without reperfusion, and this ratio is considered to be a good indicator to assess reperfusion. The results were similar to those of previous reports. The peak in all the 17 patients with confirmed reperfusion was attained within 9 hours after onset of AMI, while only 9 of the 73 patients in the group without reperfusion had their peaks within 9 hours. The diagnostic efficiency was 94%. The authors suggested a new indicator for assess reperfusion. An increase of CK-MM3 over 10% from the first to the second hour after treatment with urokinase was found in 15 of the 17 urokinase-treated patients with reperfusion. The diagnostic efficiency was also 94%. We consider that it is an indicator as good as the peak of ratio of MM3/MM1. Furthermore, with this indicator, it is possible to assess reperfusion in two hours after treatment with urokinase.


Venereal disease.

Venereal disease. Equine venereal infections of concern in the United States include EHV-3, T. equigenitalis, P. aeruginosa, and K. pneumoniae. Stallions may also harbor EAV in the genital tract and transmit the virus to mares during coitus. With the exception of EHV-3, the stallion generally remains asymptomatic while transmitting infections to mares during breeding. Methods for diagnosis, treatment, and control of these infections are discussed.


Nonpulmonary manifestations of cytomegalovirus infection in immunocompromised patients. Nonpulmonary manifestations of cytomegalovirus (CMV) infection in immunocompromised patients include chorioretinitis, gastrointestinal infection, and central nervous system disease. Diagnosis of end organ disease, especially in the gastrointestinal tract, is best substantiated by histologic evidence of CMV inclusions. Positive cultures of CMV provide evidence for supporting infection but do not define actual end organ disease. Satisfactory treatment of the disease can be accomplished with ganciclovir or foscarnet, although these agents only suppress virus replication. In many instances, severe CMV-induced end organ disease in immunocompromised patients will progress despite treatment. In some instances, resistance to the antiviral agent is the basis for drug failure. Patients at high risk for CMV disease can be identified, and studies of prophylaxis are in progress.


Cytotoxic action of cis-unsaturated fatty acids on human cervical carcinoma (HeLa) cells: relationship to free radicals and lipid peroxidation and its modulation by calmodulin antagonists. Specific fatty acids such as linoleic acid (LA), gamma-linolenic acid (GLA), dihomo gamma linolenic acid (DGLA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) showed cytotoxicity towards human cervical (HeLa) cells in vitro. Cyclo-oxygenase inhibitor, indomethacin; lipoxygenase inhibitor, nordihydroguiaretic acid (NDGA); anti-oxidant, vitamin E; and calmodulin antagonists, trifluoperazine (TFP) and chlorpromazine (CPZ) blocked the cytotoxic action of these fatty acids. GLA-induced free radical generation and lipid peroxidation were also inhibited by indomethacin, NDGA, vitamin E, TFP and CPZ. Both indomethacin and NDGA also showed significant anti-oxidant property. These results suggest that fatty acid-induced cytotoxic action against HeLa cells is a free radical dependent process and that it can be modulated by calmodulin antagonists. These results are in contrast to those observed by us earlier with human breast cancer cells where in it was found that the tumoricidal action of fatty acids can be blocked by anti-oxidants but not by cyclo-oxygenase (CO) and lipoxygenase (LO) inhibitors. From these results it can be suggested that though free radicals are the mediators of the tumoricidal action of fatty acids, the mechanism of their production may be different in different types of tumor cells.


Morphological substrates of infantile spasms: studies based on surgically resected cerebral tissue. Extensive surgical resections of neocortical cerebral tissue (including hemispherectomies) from 13 infants and children with infantile spasms showed that 12 of 13 specimens contained either malformative and dysplastic lesions of the cortex and white matter (sometimes with associated hamartomatous proliferation of globular cells), or destructive lesions possibly acquired as a result of anoxic-ischemic injury, or a combination of the two. In brain tissue from 4 patients, coarse neuronal cytoplasmic fibrils resembling neurofibrillary tangles were seen in areas of dysplastic brain on silver-stained (Bielschowsky technique) sections. Immunohistochemical (immunoperoxidase) study of cortical lesions containing globular cells employing primary antibodies to glial fibrillary acidic protein and synaptophysin as markers of astrocytic and neuronal differentiation, respectively, revealed that many cells showed astrocytic and/or neuronal features, suggesting the local proliferation of primitive or multipotential neuroectodermal cells as one substrate for this seizure disorder. Morphological abnormalities of a severe degree and wide extent in the resected tissue (e.g., in one patient with hemimegalencephaly) often showed features to suggest that they may represent variants of tuberous sclerosis. These most likely result from abnormal movement and/or local proliferation of neuroectodermal precursors that have migrated from the germinal matrix to the cortical mantle. Cellular, molecular and neurophysiological study of these abnormalities is likely to yield information about basic molecular mechanisms of brain malformation and injury important in the pathogenesis of infantile spasms and other forms of focal or generalized epilepsy.


Parallel effects of arachidonic acid on insulin secretion, calmodulin-dependent protein kinase activity and protein kinase C activity in pancreatic islets. A potential role of arachidonic acid in the modulation of insulin secretion was investigated by measuring its effects on calmodulin-dependent protein kinase and protein kinase C in islet subcellular fractions. The results were interpreted in the light of arachidonic acid effects on insulin secretion from intact islets. Arachidonic acid could replace phosphatidylserine in activation of cytosolic protein kinase C (K0.5 of 10 microM) and maximum activation was observed at 50 microM arachidonate. Arachidonic acid did not affect the Ca2+ requirement of the phosphatidylserine-stimulated activity. Arachidonic acid (200 microM) inhibited (greater than 90%) calmodulin-dependent protein kinase activity (K0.5 = 50-100 microM) but modestly increased basal phosphorylation activity (no added calcium or calmodulin). Arachidonic acid inhibited glucose-sensitive insulin secretion from islets (K0.5 = 24 microM) measured in static secretion assays. Maximum inhibition (approximately 70%) was achieved at 50-100 microM arachidonic acid. Basal insulin secretion (3 mM glucose) was modestly stimulated by 100 microM arachidonic acid but in a non-saturable manner. In perifusion secretion studies, arachidonic acid (20 microM) had no effect on the first phase of glucose-induced secretion but nearly completely suppressed second phase secretion. At basal glucose (4 mM), arachidonic acid induced a modest but reproducible biphasic insulin secretion response which mimicked glucose-sensitive secretion. However, phosphorylation of an 80 kD protein substrate of protein kinase C was not increased when intact islets were incubated with arachidonic acid, suggesting that the small increases in insulin secretion seen with arachidonic acid were not mediated by protein kinase C. These data suggest that arachidonic acid generated by exposure of islets to glucose may influence insulin secretion by inhibiting the activity of calmodulin-dependent protein kinase but probably has little effect on protein kinase C activity.


Buffering of calcium in the vicinity of a channel pore. The function of calcium entry or release channels is often modulated by the cytosolic free calcium concentration. When such channels are studied in isolation, calcium buffer solutions are usually used to control the free calcium at the cytosolic face of the channel. Such solutions are generally formulated on the basis of equilibrium considerations. We calculate the gradient of [Ca2+] in the vicinity of a channel pore, in the presence of such buffers. We find that the effective degree of buffering near the pore is markedly affected by kinetic considerations. Commonly used EGTA solutions are completely ineffective in buffering [Ca2+] within macromolecular distances of the pore. In order to achieve useful buffering, the fastest buffers (e.g. BAPTA derivatives) must be used, in concentrations very much higher than those conventionally employed. Because of the diffusion limit on the maximum rate of binding of calcium to the buffer ligand, it is physically impossible to achieve good control of [Ca2+] at cytosolic levels at distances of less than a few nm from a pore conducting pico-ampere calcium current.


Effect of isotypic and allotypic variations of MHC class II molecules on staphylococcal enterotoxin presentation to murine T cells. Staphylococcal enterotoxins (SE) are known to be potent T cell activators, stimulating +/- proliferation and lymphokine production. These toxins have recently have been termed "superantigens" because of their ability to bind directly to class II molecules forming a ligand that interacts with particular V beta gene elements within the TCR complex. This interaction between SE and MHC class II molecules plays a central role in toxin-induced mitogenesis. In the present study we have examined the effect of polymorphism on the ability of MHC class II molecules to bind and present SE. Through the use of H-2 congenic mouse strains, it was possible to look directly at haplotype differences within the MHC and their effect on SE presentation to a panel of responsive V beta-bearing T cells. The results demonstrate that toxin presentation by class II-bearing accessory cells to murine T cells is greatly affected by polymorphisms within the H-2 complex. Toxin-pulsed accessory cells obtained from mice of an H-2k and H-2u haplotype were found to be less efficient in activating a variety of T cell clones and hybridomas. However, one T cell clone responded similarly to the enterotoxins presented on all H-2 haplotypes, suggesting that differences in responses of T cells are not simply a function of the degree of binding of these toxins to various class II molecules. Neutralization analysis with monoclonal anti-class II antibodies demonstrates that both I-A and I-E molecules play a significant role in SEA and SEB presentation to murine T cells. These results suggest that the differential activation of T cells by a particular enterotoxin may reflect a difference in recognition of an SE:class II ligand by a surface T cell receptor complex.