Cytochrome aa3 depletion is the cause of the deficient mitochondrial respiration induced by chronic valproate administration. Liver mitochondria from rats fed 1% (w/w) valproic acid for 75 days displayed an approximate 30% decrease in coupled respiration rates with substrates entering the respiratory chain at complexes I and II. Uncoupling the respiration from proton-pumping, or measuring the respiration without complex IV removed this inhibition. The treatment induced a loss of activity of cytochrome oxidase consistent with a decrease in the mitochondrial content of cytochrome aa3. The inhibition induced by long lasting administration of valproate is apparently located at the site of the proton-pumping activity of complex IV. Furthermore, the capacity of electron transport through complex IV, being far in excess of that required for normal functioning in coupled mitochondria, seems to be controlled by the coupling to proton-pumping in which cytochrome aa3 appears to play a major role.


Comparison of opioid properties between D-Arg-containing dipeptides and tetrapeptides. Since the D-Arg-containing dipeptides, H-Tyr-D-Arg-OMe (TDA) and H-Tyr(Et)-D-Arg-OMe, and D-Arg2-substituted dermorphin N-terminal tetrapeptide analogues, H-Tyr-D-Arg-Phe-Gly-OEt (TDAPG) and H-Tyr(Et)-D-Arg-Phe-Gly-OEt gave different pharmacological responses in vivo, opioid interaction and structure-activity relationships have been investigated in vitro. In the isolated guinea-pig ileum assay, the tetrapeptides were potently inhibitory, their activity markedly exceeding that of the dipeptides. In particular, the first tetrapeptide had twice the activity of morphine, while the potency of the dipeptides was less than one twentieth that of morphine. Also in the opioid receptor binding assay, tetrapeptides had a higher affinity than the dipeptides. IC50 values of tetrapeptides were 8.46 and 23.7 nM, respectively, which were lower than that of morphine. Ethylation of the Tyr residue of TDA much increased the opioid activity whereas that of TDAPG greatly decreased it. All peptides used were extremely stable to aminopeptidase-M and carboxypeptidase-Y and had an inhibitory effect on enkephalin (EK)-degrading enzymes. From these results, it appears that the effects of the tetrapeptides are due mainly to specific interaction with opioid receptors, whereas the dipeptides do not act specifically on the opioid receptors, but are involved in non-opioid mechanisms. The resistance to enzymes and inhibitory effect of the peptides used on the EK-degrading enzymes may also account for their potent and long-lasting opioid-like activities.


Glucocorticoid-mediated potentiation of P450 induction in primary culture of rainbow trout hepatocytes. Induction of 7-ethoxyresorufin O-deethylase activity (a cytochrome P450IA-dependent activity) by beta-naphthoflavone (0.36 microM) is increased 2-3-fold by dexamethasone or cortisol (10(-9)-10(-7) M) in rainbow trout hepatocyte cultures. This potentiation does not seem to be a time-dependent process and could be a classical glucocorticoid receptor-mediated event resulting in enhanced transcriptional activation of the CYP1A, as previously shown in mammals. Since glucocorticoid levels can increase in fish exposed to pollutants, such steroids may interfere with the induction response to xenobiotics.


Mitochondrial complex I and II activities of lymphocytes and platelets in Parkinson's disease. Mitochondrial Complex I deficiencies have been described not only in the brain but also in the skeletal muscle and platelets in Parkinson's disease (PD). We report activities of Complex I, II, III, and IV in lymphocytes and platelets in 20 patients with PD and age-matched controls. A small but a significant decrease in the platelet Complex I activity was found in PD (9.14 +/- 1.86 units/mg protein) compared with that in the control (12.37 +/- 2.66 units/mg protein) (P = 0.0002). The lymphocyte Complex I activity was about the same between PD and the control. The activity of Complex II was slightly diminished in both platelets and lymphocytes in PD. Rather small decrease in the platelet Complex I activity in PD may be clinically non-significant. But it may indicate the presence of a qualitatively similar abnormality in platelets as in the nigro-striatal neurons. The cause for decrease in the Complex II activity is unknown at this moment. Further studies seem necessary.


Neurochemical markers in the cerebrospinal fluid of patients with Alzheimer's disease, Parkinson's disease and amyotrophic lateral sclerosis and normal controls. Several neurotransmitter markers were investigated in the cerebrospinal fluid (CSF) from patients with Alzheimer's disease (AD) (n = 27), Parkinson's disease (PD) (n = 35) and ALS (n = 26) and from control subjects (n = 34) to compare the possible alterations in the biochemical profiles of these different neurodegenerative diseases. The main proportion of the patients represented an early phase of the illness at the time of the diagnosis. Correlations of the degree of dementia and the stage of the disease with CSF measures were evaluated. The CSF levels of somatostatin like-immunoreactivity (SLI) were significantly reduced in AD patients when compared with those of normals and ALS patients. The CSF concentrations of homovanillic acid (HVA) were significantly decreased for PD patients and the decrease focused on the non-demented patients. A trend of decreasing HVA values towards the most advanced stage of Parkinson's disease assessed by Webster's scale was also displayed. The content of 3-methoxy-4-hydroxyphenylglycol (MHPG) in the CSF was higher for ALS patients than for other groups. The lowest 5-hydroxy-indoleacetic acid (5HIAA) levels were observed in the PD group and the lowest acetylcholinesterase (AChE) activities were found in the PD patients with the most severe disease. Changes in CSF measures were too subtle to be beneficial for diagnostic purposes, but adequate for reflecting the different neurochemical profiles of these three degenerative neurological disorders.


Amiloride antagonizes beta-adrenergic stimulation of cAMP synthesis and Cl- secretion in human tracheal epithelial cells. Amiloride, a potent blocker of the sodium channel in airway epithelium, has been administered by aerosol as a therapeutic agent for cystic fibrosis. Because amiloride in high concentration has been reported to interfere with cell functions, including adrenergic responses, we tested the ability of amiloride to inhibit beta-adrenergic responses in human tracheal epithelial cells. Amiloride (10(-4) M), applied from the basolateral surface of a cell monolayer, inhibited the changes in transepithelial potential and short circuit current to isoproterenol (10(-6) M). The stimulation of cyclic adenosine monophosphate (cAMP) synthesis by isoproterenol was inhibited in dose-dependent fashion by amiloride (P = 0.007 by multivariate ANOVA with multiple samples correction). Amiloride did not affect baseline transepithelial potential, short circuit current, basal cAMP levels, cAMP response to prostaglandin E2, or basal adenylate cyclase activity measured directly in membrane preparations. Therefore, it is unlikely that amiloride exerts a nonspecific toxic effect on adenylate cyclase, receptor-cyclase coupling, or substrate or cofactor supply. The binding of [125I]iodocyanopindolol (ICYP), a beta-adrenergic receptor antagonist, to membranes from human tracheal epithelial cells could be displaced by amiloride with IC50 = 410 microM; displacement was 70% at 10(-3) M amiloride. These data are most consistent with the hypothesis that amiloride inhibits beta-adrenergic responses in airway epithelial cells by occupying beta-adrenergic receptor sites. Therapeutic administration of amiloride should take into account its affinity for adrenergic receptors.


Prevalence of HTLV infection in the Dominican Republic: association with neurological disease. The presence of the human T-cell leukemia virus (HTLV) in Dominican blood donors and patients with tropical spastic paraparesis (TSP) was first detected in 1987. To define further the seroprevalence in the country, nearly 4,000 samples from high- and low-risk populations, as well as patients with neurological disease and with leukemia or lymphoma were tested for HTLV antibodies. A 1-2% seropositivity rate was found among the low-risk population, a 2-5% in the high-risk, and at least 87% in those with TSP. A few patients with malignancy also had antibodies to HTLV. An increase in seropositivity with age and a predominance of female seropositive individuals were found. Infectious virus was isolated from TSP patients, prostitutes, and family members of index patients. These data indicate the substantial level of HTLV infection in another Caribbean country and its relation to neurologic disease.


Variability of onset times within and among relaxant regimens. To evaluate the consistency of times to 95% twitch height depression (T95%) in groups of patients receiving identical induction and relaxant regimens. Prospective, noncontrolled, blinded study. Ambulatory surgical unit at a university medical center. Seventy-five ASA physical status I and II patients undergoing general endotracheal anesthesia. Patients received succinylcholine 1.5 mg/kg or a nondepolarizing regimen with doses ranging from approximately 1.5 to 6 times the ED95, with or without a priming dose. For each of the eight relaxant regimens used in five or more patients, the intraregimen variability of T95% (at the adductor pollicis muscle upon ulnar stimulation at 0.1 Hz) was expressed as SD and range, and the individual data points were displayed. There was wide intraregimen variability. For each regimen, the slowest T95% was at least 73% longer than the fastest T95%. For the 16 patients receiving a priming dose plus an intubating dose 5 or more times the ED95, the median T95% was 95 seconds; however, T95% was beyond 120 seconds in 5 of the 16 cases. The wide variability in onset times among subjects receiving the same regimen indicates that monitoring of neuromuscular response, preferably to a relatively slow rate of neurostimulation, is essential if one elects to use moderate to high doses of atracurium and/or vecuronium for rapid-sequence induction in a patient in whom movement or coughing is unacceptable. Since onset times were not symmetrical about the mean, the magnitude and frequency of unacceptable onset times would not be fully appreciated unless the individual data points were displayed. Such information may be critical when reporting the suitability of a neuromuscular blocking drug for rapid intubation.


HIV-1 in blood monocytes: frequency of detection of proviral DNA using PCR and comparison with the total CD4 count. In vivo infection of monocytes/macrophages by the human immunodeficiency virus (HIV) has been investigated in many studies since these cells were suggested to provide a reservoir for the virus. In this study, we wanted to find out whether HIV provirus could be detected in circulating monocytes and whether it could be compared with the provirus found in T lymphocytes (T-Ly). Twenty-one seropositive subjects were studied. The amplification method (PCR) was used with three different primer pairs (in gag, env, and long terminal repeat regions of the viral genome) to detect the HIV-1 genome in monocytes and T-Ly separated by an immunomagnetic isolation technique. Of 21 monocyte samples, 13 (61.9%) were positive with at least one primer pair. Furthermore, the provirus harboured in 9 of those 13 monocyte-positive samples differed, with respect to pattern of primer response, from the provirus found in T-Ly. When comparing primer responses of monocytes and T-Ly, most of the differences were found to have occurred with the env primers (8 of 9 cases). Dilution experiments with the 8 E5 cell line revealed that 9 of 12 T-Ly contained 15-150 HIV DNA copies per 150,000 cells while 8 of 11 positive monocytes contained less than 15 copies. However, monocyte samples from two asymptomatic individuals and an AIDS patient showed high levels of HIV DNA, comparable to those obtained in T-Ly. Finally, it was also found that the monocyte-positive subjects were more immunosuppressed than the negative ones, as shown by the total CD4 count of both groups (means of 269 T4/mm3 and 573 T4/mm3, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)


Comparative effects of desflurane and isoflurane on vecuronium-induced neuromuscular blockade. To evaluate the neuromuscular effects of a nondepolarizing muscle relaxant (vecuronium) during anesthesia with equipotent concentrations of either desflurane or isoflurane. Randomized open study comparing effects of desflurane and isoflurane on vecuronium-induced neuromuscular blockade. University-affiliated medical center. Forty-five healthy adults undergoing elective surgical procedures randomly assigned to receive either desflurane, nitrous oxide (N2O), and vecuronium or isoflurane, N2O, and vecuronium for maintenance of general anesthesia. Following a standardized induction sequence, patients receiving either desflurane and N2O or isoflurane and N2O were administered bolus doses of vecuronium equal to 0.01, 0.02, or 0.03 mg/kg intravenously (IV) during the maintenance period. Neuromuscular transmission was measured using a Relaxograph monitor. Vecuronium produced similar depression of neuromuscular function at equipotent (50% of the minimum alveolar concentration) end-tidal concentrations of isoflurane 0.6% and desflurane 3.0%. Following administration of vecuronium 0.01 to 0.03 mg/kg IV, onset times (3.4 +/- 0.4 minutes to 3.2 +/- 0.4 minutes and 3.2 +/- 0.5 minutes to 3.0 +/- 0.6 minutes), maximum T1 twitch depression (80% +/- 10% to 95% +/- 9% and 81% +/- 9% to 97% +/- 10%), clinical duration of blockade (12 +/- 5 minutes to 20 +/- 8 minutes and 10 +/- 5 minutes to 19 +/- 17 minutes), and T1 recovery times (10 +/- 3 minutes to 12 +/- 6 minutes and 10 +/- 3 minutes to 12 +/- 4 minutes) were similar in the isoflurane and desflurane treatment groups, respectively (means +/- SD). Vecuronium has similar neuromuscular effects when administered in the presence of desflurane 3% and isoflurane 0.6%.


Dexmedetomidine attenuates sympathoadrenal responses to tracheal intubation and reduces the need for thiopentone and peroperative fentanyl. The effects of the new, highly selective alpha 2-adrenergic agonist, dexmedetomidine, were studied in a randomized, placebo-controlled, double-blind trial in 24 ASA I patients. Dexmedetomidine 0.6 micrograms kg-1 or saline was given i.v. 10 min before induction of anaesthesia. The required dose of thiopentone was significantly (P less than 0.001) smaller in the dexmedetomidine group (mean 4.4 (sd 0.9) mg kg-1) than in the control group (6.9 (1.6) mg kg-1), and the drug attenuated the cardiovascular responses to laryngoscopy and tracheal intubation. The concentration of noradrenaline in mixed venous plasma was smaller in the dexmedetomidine group during all phases of induction (P less than 0.01). During surgery, fentanyl was required in a dose of 0.5 (0.6) mg kg-1 and 2.8 (2.6) mg kg-1 in the dexmedetomidine and control groups, respectively (P less than 0.001). During 2 h postoperative follow-up, oxycodone 0.06 (0.06) mg kg-1 and 0.16 (0.1) mg kg-1 (P less than 0.05) was given to the two groups respectively.


The role of histamine in the cardiovascular effects of atracurium. We have investigated the effect of a bolus injection of atracurium 0.6 mg kg-1 on the cardiovascular system in 16 patients undergoing aortocoronary bypass surgery. H1- and H2-receptor antagonists were administered to eight patients before the neuromuscular blocker. A standard anaesthetic was used comprising fentanyl, flunitrazepam, etomidate and enflurane. After administration of atracurium, haemodynamic changes and plasma histamine concentrations were measured at frequent intervals. In the first group, who received only atracurium, a brief but marked decrease in SVR and MAP occurred, accompanied by an increase in Cl, together with a marked increase in plasma concentration of histamine. In the second group, who received H1- and H2-receptor block, there was no decrease in MAP and only a small increase in plasma histamine concentration. However, there were significant changes in SVR and Cl similar to those in atracurium group.


Comparison of the train-of-four fade profiles produced by vecuronium and atracurium. In this double-blind study, we have allocated randomly 40 ASA I-III patients to one of four groups. After a standard anaesthetic induction, patients received vecuronium 0.08 mg kg-1 or 0.10 mg kg-1, or atracurium 0.4 mg kg-1 or 0.5 mg kg-1. Using an electromyogram (Datex Relaxograph) the train-of-four (TOF) response was measured during onset of and recovery from neuromuscular block. A greater degree of fade of TOF was observed with atracurium during onset of neuromuscular block than with equivalent doses of vecuronium. During recovery of neuromuscular transmission, vecuronium was associated with more fade than atracurium. The differences in the TOF profiles of these two drugs may be important when judging the adequacy of antagonism of neuromuscular block using the TOF response.


T4+ cell numbers are correlated with plasma glutamate and cystine levels: association of hyperglutamataemia with immunodeficiency in diseases with different aetiologies. Human immunodeficiency virus type 1 (HIV-1) seropositive individuals suffer from a depletion of T4+ T cells and have elevated plasma glutamate levels. Glutamate is also elevated in cancer patients, and several authors have shown that elevated extracellular glutamate levels inhibit competitively the membrane transport of cystine and cause a decrease of intracellular cystine. We, therefore, tested the hypothesis that high glutamate and/or low cystine levels may generally be associated with low lymphocyte reactivity or low T4+ counts. In three independent studies we tested (i) serum amino acid levels (AAL) versus T4+ counts in healthy individuals, (ii) plasma AAL versus lymphocyte responses in healthy individuals, and (iii) plasma AAL versus T4+ counts in HIV-1 seropositive individuals. When the individuals in each study were divided into four subgroups as defined by median glutamate and cystine levels, the results showed that persons with a combination of low glutamate and high cystine level (LGHC subgroups) had the highest mean T4+ count or highest lymphocyte reactivity. Moreover, the LGHC subgroup in a study of lung cancer patients had a much longer mean survival time than the other three subgroups. In HIV-1 infected patients, hyperglutamataemia is associated with hypocystinaemia and hypocysteinaemia. Azidodeoxythymidine (AZT) treated HIV patients had, on average, lower glutamate levels than patients without AZT.(ABSTRACT TRUNCATED AT 250 WORDS)


Folding of intestinal brush border enzymes. Evidence that high-mannose glycosylation is an essential early event. A polyvalent antiserum which precipitates the native, folded, but not the denatured molecular forms of pig intestinal aminopeptidase N (EC 3.4.11.2) and sucrase-isomaltase (EC 3.2.1.48, EC 3.2.1.10) was used to determine the kinetics of polypeptide folding of the two newly synthesized brush border enzymes. In pulse-labeled mucosal explants, complete synthesis of the polypeptide chains of aminopeptidase N and sucrase-isomaltase required about 2 and 4 min, respectively, whereas maximal antiserum precipitation was acquired with half-times of 4-5 and 8 min, respectively. Fructose, which induces a defective cotranslational high-mannose glycosylation, increased the half-time of polypeptide folding to about 12 min for aminopeptidase N as well as for sucrase-isomaltase. Short-pulse experiments suggested that fructose exerts its effect by slowing the rate of glycosylation, making this partially a posttranslational process. In the presence of fructose, not only the malglycosylated forms but also the electrophoretically normal, high-mannose-glycosylated form of the brush border enzymes were retained in the endoplasmic reticulum and proteolytically degraded. The results obtained demonstrate an intimate interrelationship between glycosylation and polypeptide folding in the synthesis of membrane glycoproteins and, more specifically, indicate that the timing of these two early biosynthetic events is essential for correct polypeptide folding.


The role of tryptophan in structural and functional properties of equinatoxin II. A pore-forming, cytolytic and lethal polypeptide, equinatoxin II, from the sea anemone Actinia equina, was subjected to oxidation with N-bromosuccinimide to study the role of five present tryptophan residues in structure-function relationships. In the folded toxin molecule, 1-2 tryptophan residues were readily susceptible to oxidation with N-bromosuccinimide, whereas modification of a single residue resulted in complete impairment of the toxin lethal and hemolytic activities as well as the ability of an oxidized toxin to precipitate with serum lipoproteins. CD and fluorescence spectra indicated a slight alteration of a toxin secondary structure following N-bromosuccinimide treatment. Incubation with sphingomyelin of the toxin prior to oxidation did not prevent subsequent modification with N-bromosuccinimide and loss of its activities, indicating that the modified tryptophan residue is not directly involved in toxin binding and insertion into lipid membranes. It was concluded that the modified tryptophan residue is essential for the structure of equinatoxin II.


The role of lysine, histidine and carboxyl residues in biological activity of equinatoxin II, a pore forming polypeptide from the sea anemone Actinia equina L. Equinatoxin II, a pore forming polypeptide from the sea anemone Actinia equina L. was subjected to chemical modifications with group specific reagents. Lysine residues were modified with pyridoxal-5'-phosphate, histidine residues with diethyl pyrocarbonate and carboxyl groups with the use of a water soluble carbodiimide. Modification of charged residues had no significant influence on the toxin interaction with serum lipoproteins. Lysine 5'-phosphopyridoxylated and histidine carbethoxylated derivatives of the toxin retained lethal and hemolytic activities, but the pH profile of hemolytic activity of 5'-phospho-pyridoxylequinatoxin II was markedly altered. Modification of the toxin carboxyl groups impaired both hemolytic and lethal activities, the latter, however, to the greater extent.


Conformational integrity of a recombinant toxoid of Pseudomonas aeruginosa exotoxin A containing a deletion of glutamic acid-553. A mutant form of Pseudomonas aeruginosa exotoxin A (ETA) carrying a deletion of glutamic acid-553, an important active-site residue, was expressed in an ETA-negative strain of P. aeruginosa and shown to be exported from the cells as efficiently as wild-type ETA. The mutant protein, purified from the culture medium, was devoid of ADP-ribosyltransferase activity. Protein conformation was barely perturbed by the deletion, as determined by a number of measures, including affinity for substrate NAD, proteinase sensitivity, absorbance and fluorescence spectroscopy, and differential scanning calorimetry. The conformational integrity and stability of the mutant toxin are consistent with potential use of the protein in vaccines or as a carrier in preparing conjugate vaccines.


Do antiarrhythmic doses of magnesium potentiate vecuronium? The purpose of this study was to investigate whether magnesium sulfate (MgSO4) at a dose commonly used to treat arrhythmias potentiates vecuronium. After Institutional Review Board approval, 20 randomly assigned, consenting patients received a bolus of either MgSO4 (30 mg/kg) or placebo in a blinded fashion. Immediately after receiving the bolus of either MgSO4 or placebo, the study patients were taken to the operating room (OR) and anesthetized. The ED95 of vecuronium was determined in both groups by administering 10 micrograms/kg boluses of vecuronium until 95% twitch depression was measured. Delay to 25% twitch height recovery (indicating that the neuromuscular block could be reversed for extubation) was measured in 10 patients. Ultrafilterable magnesium levels were measured in a total of 13 patients. Magnesium levels were drawn before the magnesium/placebo bolus, 5 minutes after bolus, and 30 minutes after bolus. The data did not demonstrate any relationship between the use of MgSO4 at 30 mg/kg doses and either the ED95 or the duration of vecuronium. A 30 mg/kg bolus of MgSO4 roughly doubled ultrafilterable magnesium levels from baseline. The limited sample size precluded making any firm conclusions from this data. The data trend suggested that antiarrhythmic doses of MgSO4 may not potentiate vecuronium.


The pattern of expression of the chicken homolog of HOX1I in the developing limb suggests a possible role in the ectodermal inhibition of chondrogenesis. Homeobox-containing genes have been implicated in a variety of patterning events during vertebrate limb development. In an attempt to isolate cDNAs corresponding to 5' members of the chicken HOX 4 cluster of homeobox-containing genes, a cDNA library constructed from mRNAs expressed during early stages of chick limb development was screened with probes generated by the polymerase chain reaction (PCR) using oligonucleotide primers corresponding to sequences in the homeoboxes of the human HOX4C and HOX4F genes, the human homologs of Hox-4.4 and Hox-4.6. This screening resulted in the isolation of full length cDNAs for the chicken homolog of HOX4F (cognate of mouse Hox-4.6), which we have termed GHox-4.6, and the chicken homolog of human HOX1I, which we have named GHox-1i, a paralog of Hox-4.6 in the HOX 1 cluster. The homeodomains encoded by GHox-4.6 and GHox-1i differ by only three amino acids, and the two proteins show extensive similarity along their entire lengths. Despite their sequence similarity, in situ hybridization analysis has revealed that GHox-4.6 and GHox-1i exhibit strikingly different spatial patterns of expression during embryonic chick limb development. At early stages of limb development (stages 20-22), GHox-4.6 transcripts are present in high amounts throughout the posterior half of the limb mesoderm and are absent from the anterior half of the mesoderm, an expression pattern consistent with the possible involvement of GHox-4.6 in the specification of posterior positional identity. In contrast, GHox-1i exhibits no distinct anterior-posterior polarity of expression at stage 22, but rather is expressed in high amounts throughout the mesenchyme of the limb bud. At later stages of development (stage 25), GHox-1i continues to be expressed in high amounts throughout the undifferentiated mesenchyme subjacent to the apical ectodermal ridge, and, in addition, is expressed in the mesodermal cells in the proximal peripheral regions of the limb bud subjacent to the ectoderm which are differentiating into nonchondrogenic lineages. Conversely, little or no expression of GHox-1i is detectable in the proximal central core of the limb bud where chondrogenic differentiation is occurring. Thus, GHox-1i is expressed by the undifferentiated subridge mesenchymal cells and proximal peripheral mesenchymal cells of the limb bud that are being inhibited from undergoing chondrogenesis by the apical ectodermal ridge and nonridge ectoderm.(ABSTRACT TRUNCATED AT 400 WORDS)


Tardive dyskinesia and type II schizophrenia. Cognitive impairment, negative and positive symptoms, primitive release reflexes, and age/temporal disorientation were assessed in 20 male patients meeting the DSM-III-R criteria for chronic schizophrenia and Schooler & Kane's criteria for TD. The control group comprised 20 age-matched male chronic schizophrenic patients without TD. Significant associations were found between TD, cognitive impairment, some negative symptoms, and formal thought disorder. These associations were independent of other illness and treatment variables. The severity of TD correlated significantly with that of cognitive impairment.


The antinociceptive activity of excitatory amino acids in the rat brainstem: an anatomical and pharmacological analysis. Rats were stereotaxically implanted with microinjection cannulae aimed at sites ranging caudally from the lower medulla and rostrally to the diencephalon and received microinjections of the excitatory amino acid: L-glutamate 30 nmol/0.5 microliters. The subsequent spontaneous behavioral response and the effect on the thermal noxious-evoked tail flick (TF) and hot plate (HP) responses was recorded. From 331 brain sites mapped with glutamate, an elevation of tail flick and hot plate response latencies was observed in 59 cases and in 34 of these sites the antinociceptive activity was preceded by a shortlasting aversion characterized by vocalization and running. The glutamate-sensitive sites at which TF and HP response latencies were elevated were exclusively distributed in the medullary reticular formation (MRF) and the mesencephalic periaqueductal gray matter (PAG). The aversive and antinociceptive activity of glutamate was dose-dependent and mimicked by the excitatory amino acid (EAA) receptor agonists N-methyl-D-aspartate + (NMDA) kainate and less so quisqualate. The EAA receptor antagonists MK-801 and AP-5, but not glutamyl-amino-methyl-sulfonic acid, antagonized in a dose-dependent fashion both the aversive and antinociceptive responses evoked from the PAG. It is suggested that NMDA receptor-linked neurons in the PAG activate both nociceptive and antinociceptive systems.


Brain glutamine synthetase increases following cerebral ischemia in the rat. Changes in astrocyte glutamine synthetase (GS) in postischemic rat brain were evaluated and correlated with regional neuronal vulnerability or resistance to ischemia. Rats subjected to 20 or 30 min of cerebral ischemia were allowed to survive for 3 or 24 h after ischemia; normal animals served as controls. Resultant neuronal necrosis was severe in the striatum by 24 h and in the CA1 region of the hippocampus at 72 h; neurons in paramedian cortex and CA3 region of the hippocampus were not permanently damaged. Glutamine synthetase (GS) immunocytochemistry was performed on vibratome sections of paraformaldehyde-fixed brains and enzyme activity was assayed in frozen samples of cerebral cortex, striatum and hippocampus. At 3 and 24 h after ischemia, GS immunoreactivity increased and was secondary to enlargement of GS-positive cell bodies and processes as well as to increased numbers of GS-positive astrocytes. Enzyme activity also increased in cortex, striatum and hippocampus at 3 and 24 h (P less than or equal to 0.03). This study shows that increase in astrocyte GS occurs rapidly after ischemia, and prior studies indicate that this increase occurs in parallel with proliferative changes in astrocyte organelles. The results also suggest that astrocyte metabolism of glutamate increases after ischemia. The increased capacity for glutamine synthetase may be important in normalizing extracellular glutamate following ischemia and protecting brain from the neurotoxic effects of this excitatory amino acid.


Evoked CA3 field potentials corresponding to both EPSPs and IPSPs in hippocampal slice. Biphasic field potentials were recorded in the CA3 distal dendritic region in response to both antidromic (fornix) and orthodromic (mossy fiber) stimulation in guinea pig hippocampal slices in vitro. The positive component (P1) corresponded to intracellularly recorded excitatory postsynaptic potentials. The negative component (N1) appears to be due to GABAA-mediated inhibitory postsynaptic potentials (IPSPs) since it corresponded to the fast IPSP recorded intracellularly, was blocked by bicuculline and penicillin and augmented by barbiturates. The amplitude of N1 and the duration of P1 are sensitive and useful measures of the GABAA-mediated IPSP.


Glutamate stimulation of tyrosine hydroxylase is mediated by NMDA receptors in the rat striatum. We have studied the action of glutamate on striatal tyrosine hydroxylase activity and determined which type of glutamate receptors are involved. Glutamate stimulated (EC50 = 4 +/- 2 microM) the activity of tyrosine hydroxylase in slices of rat neostriatum. The selective N-methyl-D-aspartate (NMDA) receptor antagonist 2-amino-5-phosphonovalerate (10 microM) blocked the stimulation; however, both the non-NMDA receptor antagonist glutamate diethyl ester (10 microM) and the general excitatory amino acid antagonist kynurenate (10 microM) had no effect. NMDA was even more potent than glutamate in stimulating tyrosine hydroxylase activity. Quisqualate (100 microM) only slightly stimulated the enzyme, and kainate had practically no effect. Omission of Mg2+ from the incubation medium potentiated the glutamate stimulation. Neither tetrodotoxin nor atropine prevented the stimulation. These results suggest that glutamate stimulates striatal tyrosine hydroxylase activity via NMDA receptors. The lack of effect of tetrodotoxin and atropine suggests that glutamate acts on NMDA receptors located on the dopaminergic nigrostriatal terminal. The stimulation may involve the entry of Ca2+ into the terminal through the NMDA receptor ionophore, since a Ca(2+)-free medium or cadmium totally blocked the stimulation of the enzyme by glutamate.


Age-related changes in the turnover rates of D1-dopamine receptors in the retina and in distinct areas of the rat brain. The steady-state density and the turnover rates of D1-dopamine receptors were investigated in the striatum, nucleus accumbens, substantia nigra, and retina of adult (3-month-old) and aged (23-month-old) rats. The turnover rates were measured by monitoring the repopulation kinetics of D1-dopamine receptors labeled with [3H]-SCH 23390 after the irreversible inactivation induced by a single dose of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ, 10 mg/kg, s.c.). In all the neural tissues examined, the repopulation of D1 dopamine receptors could be adequately described by a theoretical model that assumes a constant rate of receptor production (i.e. zero order) and a rate of degradation that is dependent on the receptor density at any time (i.e. first order). The results obtained indicate that the reduction in the density of D1-dopamine receptors in the striatum, nucleus accumbens and substantia nigra of aged rats is the result of a larger decrease in the receptor production rate (-44 to -60%) than in the receptor degradation rate (-21 to -46%). By contrast, the production rate of D1-dopamine receptors in the retina of aged rats remains unchanged, whilst the degradation rate is reduced by 25%. This results in an age-related increase in the density of D1-dopamine receptors in the rat retina.


Lack of serotonin neurotoxicity after intraraphe microinjection of (+)-3,4-methylenedioxymethamphetamine (MDMA). Systemic administration of 3,4-methylenedioxymethamphetamine (MDMA) produces depletions of serotonin (5-HT) and its primary metabolite, 5-hydroxyindoleacetic acid (5-HIAA), decreases 5-HT reuptake sites and diminishes tryptophan hydroxylase activity in various forebrain regions. MDMA has been shown to be neurotoxic to the fine fibers originating from dorsal raphe (DR) 5-HT neurons but not the beaded fibers from the median raphe (MR) nucleus. In the present experiment, MDMA was microinjected directly into the DR or MR to determine whether differential neurotoxicity developed in the DR versus MR fiber systems as measured by 5-HT levels and immunocytochemistry. Two weeks following stereotaxic injection with either vehicle or (+)MDMA (50 micrograms base in 2 microliters) into the DR or MR, rat brains were assayed for 5-HT and catecholamine content or 5-HT immunocytochemistry. HPLC analysis revealed no significant changes in monoamine or metabolite concentrations in the hippocampus and striatum of rats administered intra-DR or -MR (+)MDMA. Raphe sections stained for 5-HT also did not reveal any apparent neurotoxicity. A single cerebral injection of (+)MDMA does not produce neurotoxicity to 5-HT neuronal systems originating in the raphe, although neurotoxicity of multiple MDMA injections into these raphe nuclei cannot be ruled out.


Evidence for dorsal root projection to somatostatin-immunoreactive structures in laminae I-II of the spinal dorsal horn. In order to determine if somatostatin (SOM)-immunoreactive (I) cell bodies and processes in lamina (L) II of the rat spinal cord receive dorsal root input, the latter were anterogradely labeled by wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP). SOM-I structures were demonstrated by immunohistochemistry. Cell bodies labeled transscellularly or transsynaptically by WGA-HRP and immunohistochemically for SOM were present in L II. In addition, a L I cell was double labeled. These results suggest that some dorsal root axons innervate SOM-I neurons in L I-II of the spinal cord. In addition to confirming immunohistochemical observations in published reports, we have revealed SOM-I central terminals in the type II glomerulus. Further, a SOM-I CI-terminal, presumed to be of primary afferent origin, contacted a SOM-I dendrite in L II. Since SOM has been implicated in nociceptive function in the dorsal horn, it is possible that some of the SOM-I structures identified are involved in nociceptive processing.


Pattern of neuronal death in the rat hippocampus after status epilepticus. Relationship to calcium binding protein content and ischemic vulnerability. The pattern of hippocampal cell death has been studied following hippocampal seizure activity and status epilepticus induced by 110-min stimulation of the perforant pathway in awake rats. The order of vulnerability of principal cells in the different hippocampal subfields--as determined by silver impregnation--was found to be very similar to the pattern found in ischemia; i.e., dentate hilus greater than CA1, subiculum greater than CA3c greater than CA3a,b greater than dentate granule cells. The hilar somatostatin-containing cells were the most vulnerable cell type, whereas all other subpopulations of nonprincipal neurons--visualized by immunocytochemistry for the calcium binding proteins parvalbumin and calbindin--were remarkably resistant. Pyramidal cells in the CA3 region containing neither of the examined calcium binding proteins were more resistant to overexcitation than CA1 pyramidal cells, most of which do contain calbindin. This indicates that no simple relationship exists between vulnerability in status epilepticus and neuronal calcium binding protein content, and that local and/or systemic hypoxia during status epilepticus may be responsible for the ischemic pattern of cell death.


Protein kinase C isoforms in multidrug resistant P388/ADR cells: a possible role in daunorubicin transport. To identify the role of protein kinase C (PKC) isoforms in multidrug resistance in tumor cells, we examined the PKC isoform pattern in the multidrug resistant P388/ADR cell line and studied the effect of down regulation of PKC isoforms on intracellular daunorubicin accumulation and P-glycoprotein expression. Using monoclonal antibodies to PKC alpha, beta and gamma and flow cytometry technique we showed that P388/ADR cells overexpressed PKC alpha and beta as compared to drug sensitive P388 cells. Prolonged treatment of P388/ADR cells with phorbol myristate acetate (PMA), a procedure that is known to down regulate PKC, resulted in the down regulation of total PKC activity and the PKC beta isoform (at the protein level) that was accompanied by the correction of daunorubicin accumulation in P388/ADR cells. The level of expression of P-glycoprotein in PMA treated cells was similar to that of untreated cells. These results suggest that PKC beta regulates the drug efflux function of P-glycoprotein.


p53 mutations in human malignant gliomas: comparison of loss of heterozygosity with mutation frequency. Mutations in the p53 gene were analyzed in 40 gliomas using the single strand conformation polymorphism assay together with restriction fragment length polymorphism analysis to assess loss of heterozygosity for 17p alleles in the same tumors. Mutations occurred in 40% of the gliomas and were found in exons 4-8 of the p53 gene. G:C to T:A transversions, which occur in high frequency in some lung (greater than 50%), liver (greater than 80%), breast (30%), and esophageal cancers (25%), were noted in greater than 25% of the gliomas studied here. These transversions were clustered in exon 5 from codons 156 to 168, a region of the p53 gene not previously associated with a high frequency of mutation, and may represent a new hot spot for mutations in certain cancers. The majority of gliomas (27 of 38) analyzed here retained both 17p alleles. The frequency of p53 mutations was 37% in this group of tumors and increased to 64% in tumors with one 17p allele. Allelic loss for chromosome 17p occurred in 4 of 11 gliomas independently of mutations in the p53 gene. Absence of p53 mutations in 36% of the tumors with one 17p allele suggests that a tumor suppressor gene other than p53 may be located on chromosome 17p and involved in progression to malignancy of some gliomas.


Different pattern of chromosomal allele loss in multiple hepatocellular carcinomas as evidence of their multifocal origin. One of the most problematic aspects of surgery for hepatocellular carcinoma (HCC) is the frequent development of multiple tumors. Determination of the origin of multiple tumors, i.e., multifocal or metastatic, is important for predicting the clinical course of the disease after surgery. In order to clarify the origin of multiple tumors of HCC genetically, we examined patterns of loss of heterozygosity (LOH) on chromosome 16 for DNA isolated from 43 HCCs resected from 19 patients by analysis of restriction fragment length polymorphism. The cases were classified macro- and microscopically into 3 groups: multifocal origin; metastatic origin; and undetermined. Classification based on morphological features was shown to be well correlated with patterns of LOH in multiple tumors of HCC. Different patterns of LOH on chromosome 16 were detected in 8 of 11 patients with tumors of morphologically multifocal origin, whereas they were detected in none of 5 patients with tumors of morphologically metastatic origin. Among five patients with tumors of morphologically undetermined origin, a difference of LOH pattern among the tumors was detected in two, whereas in the other three, the pattern was identical between the tumors. A different pattern of LOH among HCCs arising in situ showed that they were composed of different clones, strongly suggesting their independent clonal origin and multifocal development. These results show that not only appropriate morphological observation but also examination of the LOH pattern on a particular chromosome is useful in diagnosis of multifocal HCC.


Persistent measles virus infection enhances major histocompatibility complex class I expression and immunogenicity of murine neuroblastoma cells. The effect of persistent measles virus infection on the expression of major histocompatibility complex (MHC) class I antigens was studied. Mouse neuroblastoma cells C1300, clone NS20Y, were persistently infected with the Edmonston strain of measles virus. The persistently infected cell line, NS20Y/MS, expressed augmented levels of both H-2Kk and H-2Dd MHC class I glycoproteins. Activation of two interferon(IFN)-induced enzymes, known to be part of the IFN system: (2'-5')oligoadenylate synthetase and double-stranded-RNA-activated protein kinase, was detected. Measles-virus-infected cells elicited cytotoxic T lymphocytes that recognized and lysed virus-infected and uninfected neuroblastoma cells in an H-2-restricted fashion. Furthermore, immunization of mice with persistently infected cells conferred resistance to tumor growth after challenge with the highly malignant NS20Y cells. The rationale for using measles virus for immunotherapy is that most patients develop lifelong immunity after recovery or vaccination from this infection. Patients developing cancer are likely to have memory cells. A secondary response induced by measles-virus-infected cells may therefore induce an efficient immune response against non-infected tumour cells.


Regression of left ventricular hypertrophy--a meta-analysis. Left ventricular hypertrophy (LVH) is an independent risk indicator of cardiovascular disease. Obtaining reversal of hypertension-induced cardiac hypertrophy seems to be a desirable objective of antihypertensive treatment. A total of 2,357 patients were included in a meta-analysis on the effect of antihypertensive pharmacological therapy on LVH. Overall left ventricular mass (LVM) was reduced by 11.9% (95% confidence interval (CI) 10.1-13.7) in parallel with a reduction of mean arterial pressure of 14.9% (CI 14.0 to 15.8). When evaluating the effect of first-line therapies on calculated LVM using the same formula for all studies, the absolute reductions in g were 44.7 (ACE-inhibitors), 22.8 (beta-blockers), 26.9 (calcium antagonists) and 21.4 (diuretics) when adjusted for differences between studies (ANCOVA). It can be concluded that effective antihypertensive therapy reduces LVM. ACE-inhibitors, beta-blockers and calcium antagonists reduce LVM by reducing wall hypertrophy, the effect of ACE-inhibitors being the most pronounced. Diuretics reduce LVM mainly through an effect on left ventricular inner diameter. How these effects affect prognosis is still an open question.


The secondary prevention of myocardial infarction by drug treatment; excluding lipid lowering agents. About 10% of survivors of an acute myocardial infarction will die in the following year. Thereafter the risk declines but reinfarction is still an important cause of mortality and morbidity. The post infarction trials have clearly shown that the best proven agents to mitigate this toll are aspirin, beta adrenoceptor blockers, and verapamil (but not other calcium blockers, except diltiazem for non Q wave infarction). In the context of hypertension treatment these post infarction trials may have important lessons for drug selection and ancillary treatment since the majority of subjects will ultimately die of ischaemic heart disease. Although the newer agents such as ACE and renin inhibitors, newer calcium channel blockers and alpha blockers have many promising properties in terms of risk factor reduction, no convincing mortality data exists; it is needed. This review will deal with the known effects (both good and bad) of antihypertensive agents and will also review other drug strategies relevant to the hypertensive patient. It will also point out large areas of ignorance.


An evaluation of the safety of the beta-modulator cicloprolol in chronic heart failure. The new beta-adrenoceptor partial agonist cicloprolol acts as a beta-agonist at normal levels and as a beta-antagonist at high levels of adrenergic discharge. Treatment with cicloprolol should protect the heart against excessive stimulation, while providing a baseline level of sympathetic drive. The clinical interest of such a profile is, however, not yet established in heart failure. Accordingly this study examined the safety of oral cicloprolol, a step necessary before undertaking efficacy comparison with other compounds recently proposed to treat heart failure. Twenty-five patients were studied. Cicloprolol was given once a day for 2 weeks in a crossover double-blind placebo-controlled design. Follow-ups were obtained at baseline and at the end of each period. At baseline all patients had clear evidence of heart failure. Cicloprolol did not affect resting heart rate and blood pressure, but it reduced significantly peak exercise heart rate and peak rate-pressure product. The effect was especially significant in patients with sinus rhythm. The drug did not induce bradycardia or arrhythmias. Resting and exercise ejection rate were not affected. Cicloprolol improved the quality of life and the work capacity of 5 of 12 patients with congestive failure due to ischemic etiology. Side effects were few and similar with placebo and cicloprolol. Thus, short-term administration of cicloprolol is safe in moderate heart failure.


Responsiveness of superficial hand veins to alpha-adrenoceptor agonists in insulin-dependent diabetic patients. 1. Diabetic autonomic neuropathy causes loss of sympathetic cardiovascular control and is associated with increased vascular sensitivity to catecholamines. Supersensitivity to catecholamines could be due to either a postsynaptic increase in vascular sensitivity or to decreased catecholamine uptake into peripheral sympathetic nerve endings. 2. To differentiate between these possible mechanisms we have measured the responsiveness in vivo to noradrenaline and phenylephrine with local infusions into peripheral veins of diabetic patients with and without symptomatic autonomic neuropathy and of healthy control subjects. The dorsal hand vein compliance technique was used. 3. Symptomatic diabetic patients required significantly lower doses of noradrenaline for half-maximal venoconstriction (ED50) (geometric mean 2.14 ng/min) than control subjects (geometric mean 6.61 ng/min, P = 0.032), but there was no difference in the results from the phenylephrine dose-response curves between the groups. There were no differences in venous responsiveness to noradrenaline or phenylephrine between the asymptomatic diabetic group and the control group. However, in the asymptomatic diabetic group, postural blood pressure change (an index of loss of sympathetic control) was correlated with the ED50 for noradrenaline (r = 0.74, P = 0.014), but not with the ED50 for phenylephrine. In the control group the ED50 values for noradrenaline and phenylephrine were correlated with each other (r = 0.81, P = 0.0005). 4. Both vasopressor drugs act on vascular alpha-adrenoceptors, but only noradrenaline is taken up into peripheral sympathetic nerve endings. Our results suggest that, in diabetic patients, vascular supersensitivity to catecholamines is primarily determined by decreased neuronal catecholamine uptake. A postsynaptic increase in vascular alpha-adrenoceptor stimulation does not appear to be prominent in diabetic autonomic neuropathy.


Formation of prostacyclin during administration of beta 1-selective and non-selective beta-adrenergic antagonists in healthy humans. Vascular formation of prostacyclin is increased by propranolol in patients with essential hypertension. However, the possible effect of beta-adrenoceptor blocking drugs in healthy subjects is, however, not known. We studied this issue by analysis of the urinary excretion of the prostacyclin metabolite, 2,3-dinor-6-keto-prostaglandin F1a during intake of a beta 1-selective (metoprolol) or a non-selective (propranolol) beta-adrenoceptor antagonist. After 14 days of metoprolol treatment (100 mg d-1) the urinary excretion of PGI-M did not differ from control (253 +/- 77 vs. 220 +/- 33 pg mg-1 creatinine, respectively). Five days of randomized cross-over treatment with propranolol (80 mg day-1) or placebo did not affect urinary PGI-M significantly either (177 +/- 11 vs. 202 +/- 11 pg mg-1 creatinine, respectively). Furthermore, a daily increasing dose of propranolol (80-480 mg) progressively lowered resting blood pressure and heart rate, but failed to influence urinary excretion of PGI-M. The data demonstrate that metoprolol and propranolol do not affect basal cardiovascular formation of prostacyclin in healthy subjects. Thus, the biosynthesis of prostacyclin does not appear to be regulated by beta-adrenoceptor activity under normal conditions.


Fulminant peptic ulcer disease in cardiac surgical patients: pathogenesis, prevention, and management. To identify pathogenetic factors associated with the development of peptic ulcers in patients following cardiac surgery and to examine the efficacy of medical and surgical therapy of peptic ulcers in this setting. Retrospective study with randomly selected case controls. University hospital referral practice. A total of 9,199 consecutive patients undergoing procedures requiring cardiopulmonary bypass between January 1, 1980 through September 30, 1988, were reviewed. Life-threatening ulcer complications were defined as hemorrhage of greater than 2 units of packed red blood cells which prompted subspecialty consultation and required a therapeutic intervention. Patients who developed life-threatening complications of peptic ulcers (32/9199, 0.35%) (group 1) were compared with 32 randomly selected patients (group 2) for differences in potential pathogenetic factors and outcome. Gastrointestinal hemorrhage, perforated ulcers, death. Patients in group 1 were significantly older than patients in group 2 (66.7 +/- 7.9 vs. 54 +/- 10 yrs, p less than .01). Complications following cardiopulmonary bypass requiring further surgery or causing prolonged hypotension were significantly more frequent in patients with ulcers than in controls (10/32 vs. 1/32, p less than .005). The mortality rate for patients in group 1 was 34.3% (11/32) compared with 0% in group 2 (p less than .001). Perioperative ulcer prophylaxis was employed with equal frequency in groups 1 and 2 and did not correlate with outcome. The development of complications of postoperative peptic ulcers following cardiac surgery correlates with age, need for reoperation, and hypoperfusion, but not with the use of prophylactic regimens to suppress acid secretion. These results suggest that impairment of gastric and duodenal mucosal defense mechanisms is a critical factor in the development of postoperative peptic ulcers.


A model for conversion from small volume nebulizer to metered dose inhaler aerosol therapy. Rationing of medical services will be necessary if we do not develop a more rational and efficient health care system. Respiratory care services are receiving emphasis as we try to curtail spiraling health care costs. In analysis of our respiratory care services, we found that small volume nebulizer (SVN) therapy was still a major portion of our workload. We instituted a protocol to convert to metered dose inhaler (MDI) therapy. All hospitalized patients, excluding those admitted to the spinal cord unit and intensive care units, with a physician's order for aerosol delivery by SVN, were evaluated by respiratory care practitioners for conversion to MDI therapy. A simple protocol for the therapist to use in this conversion was developed. All patients converted to MDI were trained in appropriate MDI use by the therapist. A three-day follow-up of each patient's compliance with proper MDI therapy was initiated. Even with a 72-h allowance for initial SVN treatment, we realized a 9,350 procedure reduction from deleted treatments and an additional 7,650 conversions to MDI. Less than 2 percent of our patients failed to make a completely successful conversion to MDI. Those patients who successfully converted to MDI resulted in reduced hospital costs of $43,758 based on excess medication, supplies, and labor costs associated with SVN treatments. We also saved 5,000 h of technician time that was used to further instruct patients in appropriate MDI therapy. Aerosol therapy by MDI is cost-effective therapy. The institution of guidelines for MDI conversion has reduced fear of failure for both clinicians and patients and illustrates the importance of patient education by qualified respiratory therapists.


[Clinical aspects, diagnosis and surgical therapy of persistent primary hyperparathyroidism]. Between January 1991 and September 1991 424 patients were operated on for hyperparathyroidism (HPT) at the Department of Surgery, University of Heidelberg. In 8.5% of cases a persistent HPT was present. Our own rate of persistence was 5.4%. Ultrasound revealed the pathology in 44.2% while highly selective venous catheterization identified the correct side in 63.4%. 70% of all patients with persistent HPT required one operative revision, the remaining 30% having up to 3 revisions. 27 patients (75%) were operated on successfully. In 12 of these patients an ectopic adenoma was found. The adenoma was not identified intraoperatively in 2 patients (6.9%). One patient sustained permanent damage to the recurrent laryngeal nerve.


Placebo controlled comparison of acute effects of ebastine and clemastine on performance and EEG. The effects of single oral doses of 10 and 20 mg ebastine were compared with placebo and 2 mg clemastine in a double-blind cross-over study in 16 healthy male volunteers. Clemastine produced the known pattern of changes, namely impairment of psychomotor performance, drowsiness, and a selective effect on cognitive processes. Earlier encoding in a perceptual stage was slowed whereas abstract classification processes were not affected. Electrophysiological measures of vigilance showed a general decrease in vigilance especially 2.5 and 4.5 h after dosing. In contrast at no time was any effect of ebastine different from that of the placebo. Ebastine 10 and 20 mg differed positively from clemastine in its effect on pursuit tracking, subjective rating of drowsiness and general discomfort. Ebastine 10 mg also differed positively from clemastine in the EEG features of vigilance. It is concluded that 10 and 20 mg ebastine were free from sedative adverse effects.


Characterization of the dopamine receptor expressed by rat glomerular mesangial cells in culture. Incubation of cultured rat glomerular mesangial cells with dopamine caused an increase in cyclic AMP formation in a concentration-dependent manner (Ka apparent 2.2 microM). The selective dopamine D1 receptor agonists, fenoldopam, SKF 38393 and (+/-)-2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene (6,7-ADTN) also produced concentration-dependent increases in cyclic AMP with mean Ka apparent values of 0.04 microM, 0.02 microM and 1.02 microM, respectively. Although fenoldopam and SKF 38393 were more potent than dopamine, they were partial agonists with efficacies, relative to dopamine, of approximately 60 and 35%, respectively. The dopamine analogue, 6,7-ADTN, in contrast, behaved as a full agonist. Dopamine-stimulated cAMP formation was inhibited in a concentration-dependent manner by the D1-selective antagonist, SCH 23390, with a Ki of 0.06 nM. In contrast, the D2-selective antagonist, domperidone, was four orders of magnitude less potent than SCH 23390, having a Ki of 2072 nM. In addition, SCH 23388, the stereoisomer of SCH 23390, was observed to be two orders of magnitude less potent than SCH 23390, indicating the stereoselective nature of the receptor. The potency series for the selective agonists and antagonists is the same as that described, using identical experimental conditions, for the D1 receptor expressed by a cell line of central origin confirming that the peripheral DA1 and the central D1 dopamine receptor are pharmacologically similar.


Frog cones as well as Müller cells have peroxisomes. We have localized D-amino acid oxidase in peroxisomes of frog retina using cerium procedures on tissue fixed in mixtures containing lower concentrations of glutaraldehyde than we had previously used in our cytochemical studies of this enzyme. We find the Müller cells of these preparations contain a more striking population of peroxisomes than had previously been thought: the D-amino acid oxidase-containing bodies are especially concentrated near the outer limiting membrane, but appreciable numbers are also found in the outer plexiform layer and near the inner limiting membrane. In addition, we find peroxisomes to be present in frog cone photoreceptors, particularly in zones near the ellipsoid. To our knowledge peroxisomes have not been described hitherto in vertebrate photoreceptors. Possible roles for the peroxisomes of the neural retina include participation in the metabolism of lipids (e.g. those of the cones' oil droplets, or of the outer segment) and involvement in oxidation of transmitter-related amino acids and of other small molecules.


Evidence for a protective action of the vigilance promoting drug modafinil on the MPTP-induced degeneration of the nigrostriatal dopamine neurons in the black mouse: an immunocytochemical and biochemical analysis. Based on the observations that the psychostimulant drug amphetamine in combination with physiotherapy can promote recovery of brain function after brain injury, we have studied the ability of the vigilance promoting drug Modafinil to counteract 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-(MPTP)-induced degeneration of the nigrostriatal dopamine (DA) neurons of the black mouse. MPTP was given s.c. in a dose of 40 mg/kg and the mice were sacrificed 2 weeks later. The effects of acute and chronic treatment with Modafinil were studied on MPTP-induced DA neurotoxicity. The substantia nigra and neostriatum were taken to both biochemical and histochemical analysis of presynaptic parameters of the nigrostriatal DA neurons, the latter in combination with image analysis. In separate experiments in rats in vivo tests for DA uptake blocking activity were made using intrastriatal microdialysis to study superfusate levels of DA and its metabolites and the 4-alpha-dimethylmetatyramine (H77/77) model to test for a possible ability of Modafinil to protect against H77/77-induced depletion of forebrain DA stores. Chronic treatment with Modafinil in doses of 10 to 100 mg/kg counteracted the MPTP-induced disappearance of nigral TH IR nerve cell body profiles and neostriatal TH IR nerve terminal profiles as evaluated after 2 weeks with image analysis. Chronic treatment with Modafinil (10-100 mg/kg) also dose-dependently counteracted the MPTP-induced disappearance of striatal DA uptake binding sites as evaluated at the same time interval. Also in the dose range 10-100 mg/kg Modafinil counteracts the MPTP-induced depletion of DA stores both in the neostriatum and the substantia nigra. In the acute experiments Modafinil (30 mg/kg) protected against the MPTP-induced depletion of striatal DA, dihydrophenylacetic acid (DOPAC) and homovanillic acid (HVA) levels both when given 15 min before, at the same time and 3 h following the MPTP injection. In the substantia nigra, however, these protective actions of Modafinil were only observed when the drug was coadministered with MPTP. Experiments with microdialysis in intact rats failed to demonstrate any increases of superfusate DA levels in neostriatum with 30 mg/kg of Modafinil. Modafinil in high doses of 2 x 50 mg/kg, however, significantly counteracted the H77/77 induced DA depletion of striatal DA stores. Thus, morphological and biochemical evidence has been obtained that Modafinil in the dose range 10-100 mg/kg protects against MPTP-induced degeneration of the nigrostriatal DA neurons of the black mouse.(ABSTRACT TRUNCATED AT 400 WORDS)


Medial vestibular nucleus in the guinea-pig: NMDA-induced oscillations. We have recently shown in vivo that N-Methyl-D-Aspartate (NMDA) receptors are present in the guinea-pig vestibular complex and demonstrated that they are involved in the regulation of the resting discharge of vestibular neurones. A parallel in vitro study has identified in the guinea-pig medial vestibular nuclei (MVN) two main neuronal cell types, A and B MVNn, differing by their intrinsic membrane properties. One subtype of B MVNn was further characterized by the presence of a low threshold calcium spike (LTS). The present study investigated in vitro the responses of these different cell types to NMDA. Both A and B MVNn were depolarized by NMDA, which also induced a decrease in membrane resistance and an increase in the spontaneous firing rate. These effects could be blocked by D-AP5, a specific antagonist of NMDA receptors. Following a 10-30 mV hyperpolarization, a long-lasting oscillatory behavior could be induced in presence of NMDA. These oscillations were however restricted to the subtype of B MVNn without LTS. The NMDA-induced oscillations were tetrodotoxine-resistant, but could be eliminated by D-AP5 or by replacing sodium with choline. Functional implications of this oscillatory behavior are discussed.


Receptor subtypes involved in callosally-induced postsynaptic potentials in rat frontal agranular cortex in vitro. A slice preparation of rat frontal agranular cortex preserving commissural inputs has been used for intracellular recording from layer V pyramidal cells, in order to characterize the synaptic potentials induced by stimulation of the corpus callosum and to reveal the subtypes of amino acid receptors involved. Stimulation of the corpus callosum induced EPSPs followed by early IPSPs with a peak latency of 30 +/- 2 ms and late IPSPs with a peak latency of 185 +/- 18 ms. Reversal potentials for early and late IPSPs were -75 +/- 5 mV (early) and -96 +/- 5 mV (late). Late IPSPs were more dependent on extracellular K+ concentration. The early IPSPs were blocked by GABAA antagonists, bicuculline and picrotoxin, whereas the late IPSPs were reduced by the GABAB antagonist, phaclofen. CNQX (6-cyano-7-nitroquinoxaline-2,3-dione), an antagonist of non-NMDA (N-methyl-D-aspartate) receptors, suppressed both EPSPs and late IPSPs at 5 microM. Early IPSPs remained at this concentration but were suppressed by 20 microM CNQX. In Mg(2+)-free solution, EPSPs were larger and more prolonged than in control solution. These enhanced EPSPs persisted after 5 to 20 microM CNQX, but were reduced in amplitude, and their onset was delayed by 3.6 +/- 0.8 ms. The remaining EPSPs were suppressed by 50 microM APV (DL-2-amino-5-phosphono-valeric acid), an antagonist of NMDA receptors. In Mg(2+)-free solution containing 5 to 20 microM CNQX, the late IPSPs were not diminished. The remaining late IPSPs were suppressed by APV or by phaclofen.(ABSTRACT TRUNCATED AT 250 WORDS)


[Electrophysiological studies on the hypothalamic GnRH pulse generator]. Gonadal function in mammals depends on gonadotropins secreted from the pituitary gland in a pulsatile manner. This pulsatility is governed by the periodic activation of the hypothalamic GnRH pulse generator. By means of multiple unit activity (MUA) recording techniques, characteristics increases in the neuronal activity, each of which is associated with the initiation of pulsatile LH secretion, have been recorded in the medial basal hypothalamus of the monkey, rat and goat. An unambiguous unitary relationship between the increased electrical activity (volley) and the LH pulse under a variety of physiological and experimental conditions indicates that the MUA volleys represent the electrical activity of the GnRH pulse generator. Hypothalamic MUA recordings provide direct access to the central component of the neuroendocrine control system which governs reproductive function.


[Relationship between types of hypertension and patterns of urinary catecholamine excretion in Sipple's syndrome]. We report here a case of Sipple's syndrome, and we also analyze the relationship between the type of hypertension and urinary excretion of catecholamines in Sipple's syndrome based on the literature in Japan. One hundred and fourteen cases of Sipple's syndrome have been reported in Japan. The hypertension of patients with Sipple's syndrome shows a ratio of fitfull hypertension to continual hypertension of 6 to 1, whereas the ratio is 1 to 1.5 in patients whose pheochromocytoma is not accompanied by Sipple's syndrome. The patients with Sipple's syndrome, being pheochromocytoma can be classified into the adrenaline (U-AD) dominant type and noradrenaline (U-NA) dominant type based on the catecholamine excretion in the urine. The U-AD predominant (U-AD/U-NA greater than 0.4) patients mostly reveal fitfull hypertension, while patients with continual hypertension hardly show U-AD predominant.


Isolation and characterization of genes encoding chaperonin 60 beta from Arabidopsis thaliana. Chaperonins (Cpn) are implicated in the folding and assembly of multimeric proteins in plastids and mitochondria of eukaryotes and in prokaryotes. Plastid Cpn is composed of two different polypeptides termed Cpn60 alpha and Cpn60 beta. We have isolated cDNA and genomic clones encoding Cpn60 beta from Arabidopsis thaliana. The steady-state level of the cpn60 beta mRNAs is higher in etiolated leaves and sucrose-treated plants as compared to control leaves. The A. thaliana cpn60 beta gene family consists of at least three different coding units. It was confirmed that Cpn beta-encoding genes have a high level of conservation among plants.


Highly inducible synthesis of heterologous proteins in epithelial cells carrying a glucocorticoid-responsive vector. A glucocorticoid-responsive vector is described which allows for the highly inducible expression of complementary DNAs (cDNAs) in stably transfected mammalian cell lines. This vector, pLK-neo, composed of a variant mouse mammary tumor virus long terminal repeat promoter, containing a hormone regulatory element, a Geneticin resistance-encoding gene in a simian virus 40 transcription unit, and a polylinker insertion site for heterologous cDNAs, was used to express the polymeric immunoglobulin (poly-Ig) receptor and the thymocyte marker, Thy-1, in Madin-Darby canine kidney (MDCK) cells and in murine fibroblast L cells. A high level of poly-Ig receptor or Thy-1 mRNA accumulation was observed in MDCK cells in response to dexamethasone with a parallel ten- to 200-fold increase in protein synthesis depending on the recombinant protein and the transfected cell clone.


Primary structure of the Aequorea victoria green-fluorescent protein. Many cnidarians utilize green-fluorescent proteins (GFPs) as energy-transfer acceptors in bioluminescence. GFPs fluoresce in vivo upon receiving energy from either a luciferase-oxyluciferin excited-state complex or a Ca(2+)-activated phosphoprotein. These highly fluorescent proteins are unique due to the chemical nature of their chromophore, which is comprised of modified amino acid (aa) residues within the polypeptide. This report describes the cloning and sequencing of both cDNA and genomic clones of GFP from the cnidarian, Aequorea victoria. The gfp10 cDNA encodes a 238-aa-residue polypeptide with a calculated Mr of 26,888. Comparison of A. victoria GFP genomic clones shows three different restriction enzyme patterns which suggests that at least three different genes are present in the A. victoria population at Friday Harbor, Washington. The gfp gene encoded by the lambda GFP2 genomic clone is comprised of at least three exons spread over 2.6 kb. The nucleotide sequences of the cDNA and the gene will aid in the elucidation of structure-function relationships in this unique class of proteins.


Bismuth subsalicylate in the treatment of H2 blocker resistant duodenal ulcers: role of Helicobacter pylori. Fifty nine patients with Helicobacter pylori positive duodenal ulcers that failed to heal after a six week course of treatment with H2 blockers were randomly assigned to one of the following three regimens: (i) bismuth subsalicylate, 600 mg three times daily (n = 19), (ii) ranitidine, 300 mg at night (n = 20), (iii) bismuth subsalicylate plus ranitidine (n = 20). Cumulative ulcer healing rates after four and eight weeks respectively were as follows: bismuth subsalicylate 74% (14/19) and 95% (18/19), ranitidine 40% (8/20) and 65% (13/20), bismuth subsalicylate plus ranitidine 80% (16/20) and 95% (19/20). Bismuth subsalicylate treatment was better than ranitidine at both four and at eight weeks (p less than 0.05). The clearance rates for H pylori after four weeks were: bismuth subsubsalicylate 58%, ranitidine 0%, bismuth subsalicylate plus ranitidine 55%. After stopping bismuth therapy bacterial recrudescence frequently occurred. After bismuth treatment 86% (19/22) of ulcers had healed if H pylori had been cleared, whereas only 65% (11/17) had healed if H pylori persisted (NS). This study shows that bismuth subsalicylate is more effective in the treatment of resistant duodenal ulcers than standard dose ranitidine. It may be that suppression of H pylori by bismuth subsalicylate promotes ulcer healing.


Dietary modulation of gluten sensitivity in a naturally occurring enteropathy of Irish setter dogs. Gluten sensitivity in a naturally occurring enteropathy of Irish setter dogs, and the effects of excluding dietary cereal from birth on the subsequent response to gluten challenge were investigated. Peroral jejunal biopsy specimens were obtained at 1 year of age for morphometric and biochemical examinations, and intestinal permeability was assessed using 51Cr-ethylenediaminetetraacetic acid. Affected setters, reared on a normal wheat containing diet, exhibited partial villus atrophy, intraepithelial lymphocyte infiltration, reduced brush border alkaline phosphatase activity, and increased intestinal permeability. Gluten sensitivity was shown by introduction of a gluten free diet, which resulted in resolution of morphological and biochemical abnormalities and decreased intestinal permeability, and subsequent gluten challenge, which resulted in relapse. In contrast, littermates reared exclusively on a cereal free diet showed minimal changes when challenged with gluten, apart from intraepithelial lymphocyte infiltration. These findings document a gluten sensitive enteropathy in Irish setters and indicate that exclusion of dietary cereal from birth may modify subsequent expression of the disease.


Prophylactic effects of olsalazine v sulphasalazine during 12 months maintenance treatment of ulcerative colitis. The Danish Olsalazine Study Group. In a Danish multicentre trial we compared the relapse preventing effects of olsalazine and sulphasalazine in patients with ulcerative colitis over a 12 month treatment period. Two hundred and twenty seven patients (118 men) with at least two previous attacks of ulcerative colitis were randomly allocated according to a prearranged treatment schedule to olsalazine 500 mg bd or sulphasalazine 1 g bd in a double blind, double dummy fashion. One hundred and ninety seven patients completed the trial. The relapse rate after 12 month in the olsalazine group was 46.9% v 42.4% in the sulphasalazine group with a 95% confidence interval for the difference in proportions of -9% to 18%. Seven per cent of the patients were withdrawn from the trial because of adverse drug reactions and these were equally distributed between the two groups.


Is the intercellular adhesion molecule-1/leukocyte function associated antigen 1 pathway of leukocyte adhesion involved in the tissue damage of alcoholic hepatitis? Alcoholic hepatitis is characterised histologically by an intense inflammatory cell infiltrate made up predominantly of neutrophils but including other cell types, particularly lymphocytes. Leukocyte cytotoxicity requires cell adhesion, which is mediated via receptors on the leukocyte surface including leukocyte function associated antigen-1 (LFA-1) which binds to the ligand intercellular adhesion molecule-1 (ICAM-1) on the target cell. The distribution of ICAM-1 and LFA-1 expression in liver biopsy specimens from patients with alcoholic liver disease was examined to ascertain whether this pathway of leukocyte adhesion is involved in the tissue damage of alcoholic hepatitis. Specimens were stained for ICAM-1 and LFA-1 by a three step immunoalkaline-phosphatase method using monoclonal antibodies against ICAM-1 and LFA-1. LFA-1 staining on portal tract inflammatory cells and parenchymal inflammatory cells and ICAM-1 staining on liver components were examined. ICAM-1 expression on hepatocytes was significantly greater in alcoholic hepatitis compared with fatty liver (p less than 0.001) and normal controls (p less than 0.01). ICAM-1 expression correlated with the histological degree of hepatocellular damage (tau = 0.79; p = 0.0005) and parenchymal inflammation (tau = 0.65; p less than 0.001, and with LFA-1 expression on parenchymal leukocytes (tau = 0.63; p = 0.01). The ICAM-1/LFA-1 pathway may therefore be involved in leukocyte mediated tissue damage during alcoholic hepatitis.


NEU protein overexpression in benign, borderline, and malignant ovarian neoplasms. In situ hybridization (ISH) analysis of 24 benign, borderline, and malignant ovarian tumor specimens revealed NEU transcript expression by epithelial elements in approximately two-thirds of the samples and high-level expression in 3 grade 3 adenocarcinomas. Immunohistochemical staining (IHC) of a total of 86 specimens (including 17 of those studied by ISH) localized NEU antigen expression to epithelial cells in 36 of 86 samples with strong membrane staining observed in 12, including 1 benign, 1 borderline serous carcinoma, 3 clear cell/endometrioid carcinomas, and 7 predominantly papillary serous carcinomas with areas of clear cell/endometrioid histology. Clinical correlation of the IHC results for the 72 Stage I-IV invasively malignant neoplasms revealed no statistically significant association of the intensity of NEU IHC staining with either relapse-free or overall survival. However, more of the patients whose tumors showed strong membrane staining for NEU antigen suffered relapses of disease by 3 and 4 years than did patients whose tumors showed weak or no membrane staining. These results suggest a role for the NEU gene product in the physiology of benign ovarian surface epithelium and the neoplastic epithelium of preinvasive borderline and some invasively malignant adenocarcinomas.


Polymorphism in a T-cell receptor variable gene is associated with susceptibility to a juvenile rheumatoid arthritis subset. This report demonstrates a T-cell receptor (Tcr) restriction fragment length polymorphism, defined by a Tcrb-V6.1 gene probe and Bgl II restriction enzyme, to be absolutely correlated with allelic variation in the coding sequence of a Tcrb-V6.1 gene. A pair of non-conservative amino acid substitutions distinguish the Tcrb-V6.1 allelic variants. An association of this Tcrb-V6.1 gene allelic variant with one form of juvenile rheumatoid arthritis (JRA) was established in a cohort of 126 patients. The association was observed in patients possessing the HLA-DQA1*0101 gene. Among HLA-DQA*0101 individuals, 19 of 26 patients (73.1%) carried one particular Tcrb-V6.1 gene allele as opposed to 11 of 33 controls (33%; p less than 0.005). Haplotypes carrying this HLA gene have previously been shown to confer increased risk for progression of arthritis in JRA. This demonstration of a disease-associated Tcrb-V gene allelic variant has not, to our knowledge, been previously reported and supports the contribution of polymorphism in the Tcr variable region genomic repertoire to human autoimmune disease.


Conservation of an Actinomyces viscosus T14V type 1 fimbrial subunit homolog among divergent groups of Actinomyces spp. The type 1 fimbrial subunit gene of the human Actinomyces viscosus T14V was used as a DNA probe in Southern analyses to detect related DNA sequences in 16 of 30 strains of Actinomyces spp. under conditions of high stringency. The organisms with homology to the DNA probe included two human and six nonhuman A. viscosus, three human and three nonhuman A. naeslundii, and two A. bovis isolates. Homologous DNA sequences were not detected in strains of A. odontolyticus and A. israelii examined in this study. Northern (RNA) blot analysis revealed expression of a transcript from each of the A. viscosus and A. naeslundii strains and from one A. bovis strain that was comparable in size to that detected from A. viscosus T14V. Cell surface fimbriae were observed on a majority of the strains that expressed the transcript. Various degrees of cross-immunoreactivities between these strains and antibodies specific for type 1 fimbriae of A. viscosus T14V were also observed by colony immunoassay. Thus, the data clearly demonstrate the existence in, and expression by, divergent Actinomyces groups of genomic sequences that are closely related to the type 1 fimbriae of A. viscosus T14V.


Isolation and characterization of Actinomyces viscosus mutants defective in binding salivary proline-rich proteins. Recent studies have provided evidence for human salivary proline-rich proteins (PRPs) serving as potential receptors in the acquired pellicle for Actinomyces viscosus type 1 fimbriae. We report here the isolation of mutants derived from A. viscosus T14V-J1 which are defective in binding to PRPs partially purified from parotid gland saliva. Mutagenesis with ethyl methanesulfonate preceded enrichment for cells nonreactive with PRPs by successive adsorptions with PRP-treated latex beads. Screening was accomplished by random selection of 250 isolated colonies from each of four enrichment cycles and reaction with PRP-treated latex beads in microtiter plates. Two mutants of independent origin were examined for adherence to hydroxyapatite treated with either PRPs, proline-rich glycoproteins, deglycosylated proline-rich glycoproteins, or whole saliva. Additional surface properties that were examined included agglutination with polyclonal antisera to type 1 and type 2 fimbriae, agglutination by a monoclonal antibody to type 1 fimbriae that inhibits adherence of the parent strain to saliva-treated hydroxyapatite, the ability to bind monoclonal antibody to the type 1 fimbrial subunit, and lactose-reversible coaggregation with Streptococcus sanguis 34. Both mutants exhibited reduced binding to hydroxyapatite treated with whole saliva or salivary protein preparations but were still capable of reaction with antiserum to type 1 and type 2 fimbriae. In addition, these mutants possessed the ability to bind monoclonal antibody to the type 1 fimbrial subunit in amounts comparable to the amount bound by the parent strain but were not agglutinated by the adherence-inhibiting monoclonal antibody. When considered with previously published data, these results suggest that an adhesive molecule is probably associated with type 1 fimbriae and allows for the interaction of A. viscosus with constituents in the salivary pellicle.


Fimbriation, capsulation, and iron-scavenging systems of Klebsiella strains associated with human urinary tract infection. Thirty-two strains of Klebsiella pneumoniae and seven strains of Klebsiella oxytoca isolated from urinary tract infections in elderly adults were analyzed for capsular antigens, iron-scavenging systems, and fimbriation. All strains were capsulated. Twenty-seven different K antigens were identified among the strains, with no particular antigen dominating. All strains produced the iron-scavenging system enterochelin as analyzed by bioassay and DNA hybridization. In contrast, the aerobactin iron-sequestering system was not detected in any of the strains. All strains caused hemagglutination of tannin-treated human erythrocytes and reacted with an anti-type 3 fimbriae antiserum as well as in DNA hybridization with a type 3 fimbria-specific probe, indicating that the Klebsiella strains possessed this fimbrial type. Possession of type 1 fimbriae was analyzed by agglutination tests and by hybridization with DNA probes from two distinct Klebsiella type 1 fimbria gene clusters. Phenotypic expression of the type 1 fimbriae was found in 29 of 32 K. pneumoniae strains, whereas 30 strains reacted with either of the two type 1 fimbrial cluster DNA probes. In K. oxytoca, however, only three of seven strains expressed type 1 fimbriae and reacted with the DNA probes. The type 3 fimbriae were found to bind to a fraction of epithelial cells exfoliated in normal human urine, whereas the type 1 fimbriae bound strongly to urinary slime. No inhibitors of type 3 fimbrial binding were detected in human urine.


Identification of two porcine brush border glycoproteins that bind the K88ac adhesin of Escherichia coli and correlation of these glycoproteins with the adhesive phenotype. In this study, we identified two brush border glycoproteins (210 and 240 kDa) that bind both K88ac+ Escherichia coli and purified K88ac adhesin. The specificity of these binding glycoproteins for the K88ac adhesin was demonstrated in studies in which the binding of 35S-labeled K88ac+ E. coli and biotinylated K88ac adhesin to these glycoproteins was blocked in the presence of a 100-fold molar excess of unlabeled K88ac adhesin but not in the presence of the K99 adhesin. Pretreatment of adhesive brush borders with sodium metaperiodate destroyed both binding activities, indicating that the interaction between the K88ac adhesin and the binding glycoproteins requires the glycoprotein carbohydrate moiety. It was demonstrated previously that K88ac+ E. coli binds to adhesive brush borders but not to nonadhesive brush borders (R. Sellwood, R. A. Gibbons, G. W. Jones, and J. M. Rutter, J. Med. Microbiol. 8:405-411, 1975). In the present study, brush borders isolated from 10 different pigs were tested first for brush border adhesiveness and then for the presence of the binding glycoproteins. In all cases, the binding glycoproteins were detected only in the adhesive brush border preparations. These two binding glycoproteins may be the receptors used by K88ac+ ETEC to adhere to intestinal brush border cells. Their presence on adhesive brush borders and absence on nonadhesive brush borders may be the basis for resistance and susceptibility of pigs to K88ac+ ETEC infections.


Passive protective effect of chicken egg yolk immunoglobulins against experimental enterotoxigenic Escherichia coli infection in neonatal piglets. Passive protection of neonatal piglets against fatal enteric colibacillosis was achieved with powder preparations of specific antibodies against K88, K99, and 987P fimbrial adhesins of enterotoxigenic Escherichia coli. The antibody powders were obtained by spray drying the water-soluble protein fraction of egg yolks from immunized hens after the lipid components were precipitated with an aqueous dispersion of acrylic resins (Eudragit L30D-55; Rohm pharma). The anti-K88, -K99, and -987P antibody preparations reacted specifically against the corresponding fimbrial antigens in an enzyme-linked immunosorbent assay. The orally administered antibodies protected in a dose-dependent fashion against infection with each of the three homologous strains of E. coli in passive immunization trials with a colostrum-deprived piglet model of enterotoxigenic E. coli diarrhea. Scanning electron microscopy revealed adherence of enterotoxigenic E. coli in intestinal epithelial surfaces of control piglets, whereas in treated piglets treated with high-titer antibodies, a resistance to bacterial adhesion was observed. An enzyme immunoassay with avidin-biotin complex demonstrated specific local antibody activity in target areas of the small intestines. In vitro, E. coli K88+, K99+, and 987P+ strains adhered equally to porcine duodenal and ileal epithelial cells but failed to do so in the presence of homologous anti-fimbrial antibodies. Absorption of egg yolk antibodies with fimbrial immunosorbent removed the anti-fimbrial antibody fraction and reduced significantly the protective nature of the antibody preparation in a passive immunization experiment, suggesting that anti-fimbrial antibodies were the active components.


Nadolol to treat aggression and psychiatric symptomatology in chronic psychiatric inpatients: a double-blind, placebo-controlled study. Considerable evidence indicates that the lipophilic beta-blocker propranolol is useful in treating organically based aggression. This study looked at the efficacy of a more hydrophilic beta-blocker, nadolol, to treat aggression in chronic psychiatric inpatients. Forty-one chronic psychiatric inpatients with an average of one aggressive outburst per week (defined by the Overt Aggression Scale [OAS]) were entered into a double-blind, placebo-controlled study lasting 17 weeks. The OAS was used to track aggression on a per-incident basis, while the Brief Psychiatric Rating Scale (BPRS) and the Clinical Global Impressions scale (CGI) were used to track clinical status. Nadolol subjects showed a significant decline in frequency of aggression compared with controls (p = .026) and a significant decline in the BPRS total score (p = .007) and in the subfactors "hostility and suspicion," "negative symptoms," and "signs of hyperarousal/tension." There was no significant change in CGI "severity of illness" ratings between groups, although the nadolol group was significantly improved from baseline at every subsequent time period while the placebo group was unchanged throughout the study. Nadolol is of significant benefit in the treatment of aggression in chronic psychiatric inpatients. This drug does penetrate the brain over time, but the success of a drug whose primary locus of action is peripheral may implicate a bimodal mechanism of action, i.e., a role for the CNS and the soma in the maintenance of aggression.


A comparison of diagnostic criteria for neuroleptic malignant syndrome. A variety of diagnostic criteria for neuroleptic malignant syndrome (NMS) have been used in clinical studies of this disorder, but it is not known if different criteria consistently identify NMS. This study examines agreement between three frequently used sets of diagnostic criteria in a series of possible NMS episodes. All clinically suspected NMS episodes occurring at a large tertiary psychiatric facility during a 6-year period were evaluated by three different sets of diagnostic criteria. Agreement among these criteria was quantified statistically by means of the kappa and intraclass correlation coefficients. The NMS diagnostic criteria examined generally demonstrated only fair agreement with one another in the diagnosis of NMS. Agreement was best among these criteria when the "probable" category was employed. A complex interaction involving both definition and structure of individual diagnostic criteria and designation of criteria as major or minor appears to contribute to these findings. The published diagnostic criteria used in this study do not consistently identify NMS episodes and demonstrate different thresholds for assigning this diagnosis. These differences are not due solely to different definitions of individual criteria (e.g., fever). Possible implications of these findings for clinical practice and research are discussed.


Symptomatic predictors of ECT response in medication-nonresponsive manic patients. Relations between pretreatment symptom ratings and ECT outcome were examined in acute manic patients admitted at a state psychiatric center who were nonresponsive to medication. One or 2 days after undergoing pretreatment clinical ratings, patients were randomly assigned to intensive pharmacotherapy, right unilateral ECT, left unilateral ECT, or bilateral ECT. Patients who failed to respond to any of the first three treatment conditions were assigned to crossover bilateral ECT. Patients who failed to respond to a primary treatment condition other than bilateral ECT but refused crossover bilateral ECT and those in whom ECT was terminated due to an organic brain syndrome were not considered for this study. No psychotropic drugs were prescribed for 4-7 days before, or during, the course of ECT. Of the 24 patients who entered the study, 18 patients completed the protocol. Based on independent clinical assessments by two research psychiatrists, a favorable treatment response was defined as a complete recovery from manic episode that was sustained for 1 week during which no psychotropic medications were prescribed. ECT nonresponders were significantly more angry, irritable, and suspicious than ECT responders. Severity of mania and depression ratings at baseline were not related to ECT response. Symptoms associated with poor response to ECT may overlap with those that have been reported to predict nonresponse to treatment with lithium. Our evidence also may support the view that differential symptom patterns may predict treatment outcomes to ECT and carbamazepine, respectively, in manic patients who do not respond to treatment with lithium or neuroleptics.


Purification and determination of glutamine synthetase by high-performance immunoaffinity chromatography. High-performance immunoaffinity chromatography (HPIAC) with anti-glutamine synthetase polyclonal antibodies bound to epoxy-activated silica was used to purify and determine this enzyme from the cyanobacterium Synechocystis. A single-step HPIAC procedure with cell-free extracts yielded electroporetically homogeneous glutamine synthetase. In the determination of glutamine synthetase by HPIAC a linear response in the range 10-60 micrograms of enzyme was observed. Recoveries of 70% of the loaded enzymatic activity and 100% of protein were obtained. The determination of glutamine synthetase protein by HPIAC was compared with that obtained by rocket immunoelectrophoresis. The chromatographic method is proposed as a possible alternative to other immunochemical quantitative techniques, particularly when non-limiting amounts of samples are available.


Changes in normal glycosylation mechanisms in autoimmune rheumatic disease. To investigate potential mechanisms controlling protein glycosylation we have studied the interrelationship between lymphocytic galactosyltransferase (GTase) activity and serum agalactosylated immunoglobulin G levels (G(0)) in healthy individuals and patients with rheumatoid arthritis and non-autoimmune arthritis. In RA there was reduced GTase activity and increased G(0). A positive linear correlation between B and T cell GTase was found in all individuals. The relationship between GTase and G(0) was found to be positive and linear in the control population and negative and linear in the RA population. Sulphasalazine therapy maintained normal levels of GTase and caused a reduction in G(0) in the RA population. IgG anti-GTase antibodies (abs) were significantly increased in the RA population, whereas IgM anti-GTase abs were significantly decreased in both the RA and the non-autoimmune arthritis groups. These data describe a defect in RA lymphocytic GTase, with associated abnormal G(0) changes, which is corrected by sulphasalazine. A possible regulatory mechanism controlling galactosylation in normal cells is suggested, in which there is parallel control of B and T cell GTase. IgM anti-GTase abs may be integrated into this normal regulatory process. This is disrupted in RA, where the positive feedback between GTase and G(0) is lost and there is an associated increase in IgG anti-GTase abs, which may result from isotype switching as IgM anti-GTase abs are reduced. We suggest that these mechanisms are of relevance to the pathogenesis of RA, and that their manipulation may form part of a novel therapeutic approach.


Genetic study of a new X-linked recessive immunodeficiency syndrome. Seven forms of X-linked (XL) immunodeficiency have been described (XL agammaglobulinemia, XL severe combined immunodeficiency [SCID], Wiskott-Aldrich syndrome, XL chronic granulomatous disease, XL hyper-IgM syndrome with low IgG and IgA, and XL lymphoproliferative syndrome), and properdine deficiency. Although there are (some) phenotypic variants, diagnosis is relatively simple on the basis of clinical, immunological, and genetic characteristics. We studied a family in which several males were affected by severe infections and whose pedigree suggested recessive XL inheritance of an immunodeficiency. Immunologic and genetic studies (X inactivation patterns in females and restriction fragment length polymorphism [RFLP] segregation) were performed in order to characterize the immunodeficiency. The propositus, a 5-yr-old boy, was found to have a severe and progressive T- and B-cell functional immunodeficiency characterized by defective antigen-specific responses. No lymphocyte subsets or membrane anomalies were detected and the immunodeficiency did not correspond to usual XL forms. Studies of DNA from two of the informative females, the mother and one sister revealed nonrandom X chromosome inactivation of T cells and, partially, B cells but not PMN, a pattern similar to that observed in XL SCID carriers. RFLP studies identified a haplotype segregating with the abnormal locus that may be localized in the proximal part of the long arm of the X chromosome. We thus report the characterization of a new XL immunodeficiency that may correspond either to another XL locus or to an attenuated phenotype of XL SCID.


Genetic linkage of type VII collagen (COL7A1) to dominant dystrophic epidermolysis bullosa in families with abnormal anchoring fibrils. Epidermolysis bullosa (EB) in a group of genodermatoses characterized by the fragility of skin. Previous studies on the dystrophic (scarring) forms of EB have suggested abnormalities in anchoring fibrils, morphologically recognizable attachment structures that provide stability to the association of the cutaneous basement membrane to the underlying dermis. Since type VII collagen is the major component of the anchoring fibrils, we examined the genetic linkage of dominant dystrophic EB (EBDD) and the type VII collagen gene (COL7A1) locus, which we have recently mapped to chromosome 3p, in three large kindreds with abnormal anchoring fibrils. Strong genetic linkage of EBDD and COL7A1 loci was demonstrated with the maximum logarithm of odds (LOD) score of 8.77 at theta = 0. This linkage was further confirmed with two additional markers in this region of the short arm of chromosome 3, and these analyses allowed further refinement of the map locus of COL7A1. Since there were no recombinants between the COL7A1 and EBDD loci, our findings suggest that type VII collagen is the candidate gene that may harbor the mutations responsible for the EB phenotype in these three families.