Digital rectal examination-associated alterations in serum prostate-specific antigen. The effect of digital rectal examination (DRE) on measurement of serum prostate-specific antigen was investigated during a prostate cancer screening program of 2,736 ambulatory men. Serum samples were collected before and after DRE and values compared using the nonparametric Wilcoxon signed rank test. Small, yet statistically significant, increases were found associated with DRE. The magnitude of these increases, however, was of minor clinical importance. Patients who exhibited the largest increases with initial values greater than the upper limit of normal (4 micrograms/L) were found to have either benign prostatic hyperplasia, prostatitis, or prostatic carcinoma. The benign prostatic hyperplasia patients showed relatively low initial prostate-specific antigen values with similarly small increases related to DRE, whereas the prostatitis and cancer patients exhibited both higher initial prostate-specific antigen values and larger increases associated with DRE. Finally, patients with increases in prostate-specific antigen from less than 4 micrograms/L to greater than or equal to 4 micrograms/L comprised less than 2% of the reference range population, the majority of whom had post-DRE measurements of less than 5 micrograms/L. Thus, DRE does not appear to be a significant factor in falsely elevating prostate-specific antigen levels and should be of limited concern to the clinician obtaining serum samples after DRE.


Current issues in maternal serum alpha-fetoprotein screening. Measurement of maternal serum alpha-fetoprotein originated in the early 1970s as a means to screen for fetal neural tube defects, a relatively common and devastating class of malformations. Since that time, assay methods have improved, interpretation has been refined, follow-up testing for neural tube defects has advanced, and many other disease associations have been uncovered. It is a unique test, both in its clinical application and its laboratory implementation. The present review outlines current procedures for maternal serum alpha-fetoprotein screening and summarizes recent developments.


Eosinophilia associated with chronic pancreatitis: an analysis of 122 patients with definite chronic pancreatitis. Among 122 patients with chronic pancreatitis, marked eosinophilia (greater than 500 eosinophils/mm3 in the peripheral blood) was observed in 21 cases (17.2%). All of the affected patients were males, and there was no significant difference in the incidence of eosinophilia between patients with alcoholic and nonalcoholic pancreatitis. In the patients with eosinophilia, endocrine pancreatic function was maintained comparatively well, despite marked exocrine pancreatic dysfunction. The eosinophilia of chronic pancreatitis frequently developed in association with severe damage to neighboring organs (pleural effusion, pericarditis, and ascites), as well as in association with pancreatic pseudocyst. Our findings suggest that there is a close correlation between marked eosinophilia and severe tissue injury during acute exacerbations of chronic pancreatitis.


Effect of a high dose of ethanol on serum pancreatic enzymes in young healthy adults. Short-term effects of a high dose of ethanol on the serum activities of pancreatic enzymes were studied in young healthy adults. There were 10 males and 10 females in the study group, and two males and three females in the control group. In the study group, ethanol (2 g/kg of body weight) was given during 4 h, resulting in the blood ethanol level of 1.6 +/- 0.3 g/l at 6 h. No significant changes in serum pancreatic enzyme activities were observed in the control group during the 56-h experiment. In the study group, none of the individuals demonstrated a marked increase (double from the baseline) in serum pancreatic enzyme activities. In the male study group, the serum total amylase activity increased slightly at 6 h, but the serum pancreatic isoamylase activity remained unchanged. These data suggest that, in nonalcoholic individuals, short-term exposure to a high dose of ethanol does not induce such injury in the pancreas that would appear as a leak of enzymes from the acinar cells into the circulation.


Granulomatous gastritis and Whipple's disease. Granulomatous gastritis is an uncommon morphological diagnosis. An etiopathogenetic diagnosis can be reached only by combining the morphological examination with clinical and laboratory investigations. The diagnosis of Whipple's disease in our patient was based upon a classical clinical picture and upon the likewise classical morphological appearance of the small intestinal mucosal biopsy. To our knowledge, this case report is the first to describe granulomatous gastritis in a patient with Whipple's disease.


Maternal serum alpha-fetoprotein screening and fetal chromosome anomalies: is lowering maternal age for amniocentesis preferable? We have compared the cytogenetic abnormalities diagnosed prenatally in 1,098 patients referred for amniocentesis because of low maternal serum alpha-fetoprotein (MSAFP) to those of 445 patients whose indication was elevated MSAFP and those of 361 patients who had amniocentesis for "maternal anxiety." Autosomal trisomies, sex chromosome aberrations, and various structural rearrangements were detected in all 3 groups and actually exceeded the age-related incidence estimates. The frequency of chromosome anomalies in cases studied because of "maternal anxiety" with no prior screening was similar to that in the group referred for low MSAFP (1.38 and 1.27%, respectively). A relatively higher frequency (2.02%) was detected in the group whose indication was elevated MSAFP. Maternal serum screening is designed primarily to recalculate risk figures for Down syndrome, but not for other major chromosome abnormalities. The concept of prenatal screening for chromosome aberrations must therefore be reevaluated. We think that efforts should be directed at making amniocenteses more accessible to patients who request it. "Lowering" maternal age limits to 30 would encompass a greater proportion of pregnancies at risk and would be a step toward more effective prenatal diagnosis for chromosome abnormalities.


Depression and Parkinson's disease: a review. The purpose of this review is to provide an update of the research regarding depression in Parkinson's disease and to synthesize the information into a neurobiological model relating the structural and biochemical changes in this disorder to the behavioral manifestations. The author used a computer-based search of the literature, augmented by extensive bibliography-guided article reviews, to find information on depression and Parkinson's disease. Depression occurs in approximately 40% of patients with Parkinson's disease; depression in Parkinson's disease is distinguished from other depressive disorders by greater anxiety and less self-punitive ideation. Lower CSF levels of 5-hydroxyindoleacetic acid, a past history of depression, and greater functional disability are associated with a greater risk of depression in Parkinson's disease. Female gender, early age at onset of Parkinson's disease, and greater left brain involvement may also be risk factors. Approximately half of depressed patients with Parkinson's disease meet criteria for major depressive episodes; half have dysthymia. Depression is more common in Parkinson's disease with prominent bradykinesia and gait instability than in tremor-dominant syndromes. Depressed patients with Parkinson's disease have greater frontal lobe dysfunction and greater involvement of dopaminergic and noradrenergic systems than nondepressed patients with the disease. Mood changes in Parkinson's disease respond to treatment with conventional tricyclic antidepressants or ECT. Neurobiological investigations suggest that depression in Parkinson's disease may be mediated by dysfunction in mesocortical/prefrontal reward, motivational, and stress-response systems. Neuropsychological, metabolic, clinical, pharmacological, and anatomical studies support the involvement of frontal dopaminergic projections in patients with Parkinson's disease and depression.


Macrophage development: I. Rationale for using Griffonia simplicifolia isolectin B4 as a marker for the line. The isolectin B4 of Griffonia simplicifolia (GSA I-B4) binds to cell membrane glycoconjugates bearing terminal alpha-D-galactose, which macrophages possess. We have investigated the merits of its use as a marker for cells of this lineage when examining the early origin of macrophage populations in rat embryos, the stages and time scale of transformation from precursor forms to active, matured cells, and the response of precursors and macrophages to colony-stimulating blood factors, the last two studies conducted in organ cultures of prenatal lungs. In the present instance, GSA I-B4 was used either coupled with fluorescein (FITC) for light microscopy of living and fixed cells, or with peroxidase for light or electron microscopy. Control incubations of lung culture-derived macrophages proved that staining resulted from specific binding to galactosyl units on the cell membrane, since it was competitively inhibited by alpha-D-galactose. The lectin binds to few cells in 14-day prenatal lung explants but to a great many macrophages that subsequently develop in the cultures, indicating that it can be relied on for quantitative studies on population growth; however, it is important to provide reagents with good access to the cells. Apart from macrophages and their precursors, virtually no cells in prenatal lung cultures bind this lectin. Granulocytes of adult blood are GSA positive, but they are not yet present in 14-day prenatal explants and do not develop subsequent to culturing; hence they are not a source of confusion for experimental studies using this system. Precursors of granulocytes begin to appear in rat embryos around day 13 and have GSA-positive cell membranes, but like definitive granulocytes they also have conspicuous peroxidase-positive lysosomal granules which serve to distinguish them from early macrophages, particularly when cells are studied at an ultrastructural level. With these objections cleared away, GSA I-B4 emerges as a valuable means to mark cells of the macrophage line, mature or immature.


Effects of neuraminidase on airway reactivity in the guinea pig. We investigated the effects of neuraminidase, a viral enzyme that cleaves alpha ketosidic cell-bound sialic acids, to see if it accounts for parainfluenza and influenza virus-induced airway hyperreactivity. Accordingly, Vibrio cholerae neuraminidase was administered intratracheally in guinea pigs, and airway reactivity was assessed 3 h later. Removal of sialic acid residues was evaluated by histologic studies. Airway responsiveness was determined in anesthetized, tracheotomized, and mechanically ventilated guinea pigs by exposing them to increasing concentrations of aerosolized bronchoconstrictor agents. Respiratory system conductance was measured by the occlusion method. Neuraminidase injected intratracheally did not change airway reactivity to 10(-4) to 10(-2) M acetylcholine or 10(-4) to 2.5 x 10(-3) M histamine; nor did it prevent aerosolized albuterol from inhibiting histamine-induced bronchoconstriction. Substance P (10(-6) to 5 x 10(-5) M) had no significant bronchoconstrictor effect on guinea pigs pretreated with saline or neuraminidase. In guinea pigs pretreated with aerosols of the neutral endopeptidase inhibitor phosphoramidon (10(-4) M) before the concentration curve to aerosolized substance P was recorded, neuraminidase significantly reduced substance P-induced bronchoconstriction. When bronchoconstriction was induced by the 4-11 fragment of substance P (10(-5) to 10(-2) M), which is devoid of positive charges, it did not differ significantly in guinea pigs pretreated with saline and those pretreated with neuraminidase. These results indicate that in the guinea pig, neuraminidase injected intratracheally does not induce non-specific airway hyperreactivity and may alter the binding of substance P to its receptors.


Disposition of beta-hexachlorocyclohexane, p,p'-DDT, and trans-chlordane administered subcutaneously to monkeys (Macaca fascicularis). To evaluate skin lipid analysis for the accumulation level of environmental pollutants, the correlations between organochlorine pesticide residues in adipose tissue, blood, and skin lipids of monkeys were studied. The mixture of beta-hexachlorocyclohexane (beta-HCH), p,p'-DDT, and trans-chlordane was subcutaneously given to monkeys once weekly for 5 weeks at dose levels of 1 and 10 mg/kg. The chemicals distributed in adipose tissue, blood, and skin lipids were determined six times after the last dosing at intervals of 4 to 9 weeks. Oxychlordane and p,p'-DDE were detected in all tissues together with the administered chemicals. In blood and adipose tissue, trans-chlordane decreased rapidly and oxychlordane and p,p'-DDE increased gradually and then remained at constant levels. beta-HCH and p,p'-DDT in adipose tissue increased until the 12th week and then decreased in all animals. The correlation coefficients between blood and adipose tissue regardless of dose level and collection time for each chemical ranged from 0.83 to 0.94. Correlation coefficients between skin lipids and adipose tissue varied with the chemical, namely, 0.31, 0.72, 0.81, 0.81, and 0.83 for p,p'-DDE, trans-chlordane, p,p'-DDT, beta-HCH, and oxychlordane, respectively. The results indicated that skin lipid analysis may be useful for the evaluation of specific pollutants in the body burden.


Effect of deoxynivalenol on neurotransmitters in discrete regions of swine brain. The effect of deoxynivalenol (DON, vomitoxin) on brain amine levels was investigated in swine. DON, a trichothecene mycotoxin, causes suppression of feed intake (anorexia) in susceptible species. Following acute administration of DON to pigs (0.25 mg/kg, IV), concentrations of endogenous catecholamines norepinephrine (NE), dopamine (DA), 3,4-dihydroxyphenyl-acetic acid (DOPAC), and homovanillic acid (HVA), and the indoleamines, 5-hydroxytryptamine (5HT, serotonin) and 5-hydroxyindoleacetic acid (5HIAA) were determined in five brain regions, periodically during the 24 h post-dosing. Analysis was carried out by high performance liquid chromatography, using electrochemical detection. Effects of DON in the swine brain were transmitter, time and region-specific. It was observed that levels of the major transmitters (NE, DA and 5HT) were statistically different from controls in the hypothalamus (Hypo), frontal cortex (FCX) and cerebellum (Cb) up to 8 h post-dosing. Overall, DON administration elevated NE and depressed DA concentrations in these regions, and levels of 5HT which increased initially in Hypo (1 h), had dropped significantly below controls in both Hypo and FCX at 8 h. These alterations, however, were not indicative of known neurochemical changes associated with chemical-induced anorexia. Instead, this data suggested that the neurochemical effects of acute DON exposure might be due to peripheral toxicological events (i.e., vomiting), which overwhelmed its more subtle feed refusal activity.


Proteoglycan synthesis in human erythroleukaemia (HEL) cells. Synthesis of sulphated proteoglycans was compared in human erythroleukaemia (HEL) cells grown under control conditions and under stimulation by dimethyl sulphoxide (DMSO) and phorbol 12-myristate 13-acetate (PMA). Synthesis of [35S]sulphate-labelled proteoglycans by DMSO-treated cells was decreased by about 35% relative to controls, but synthesis of proteoglycans by PMA-treated cells increased 3-4-fold. Control and DMSO-treated cells secreted 65% of the newly synthesized proteoglycans, but PMA-treated cells secreted more than 90%. Sepharose CL-6B chromatography and SDS/PAGE suggested the presence of several proteoglycans in the cells and culture medium. The PMA-treated cells synthesized a low-Mr proteoglycan (Kav. 0.3( that was not present in controls and DMSO-treated cultures. The proteoglycans of the cells and medium from control, DMSO-treated and PMA-treated cultures could be separated into three fractions by octyl-Sepharose chromatography. The proteoglycans were resistant to trypsin but were degraded by Pronase and papain to fragments similar in size to the NaOH/NaBH4-generated glycosaminoglycans. The average chain length of the glycosaminoglycans (Kav. 0.20 on Sepharose CL-6B for controls) was decreased by DMSO (Kav. 0.25) and by PMA (Kav. 0.30-0.38). Chondroitin ABC lyase digestion of the proteoglycans from the medium of the control cultures produced two core proteins at Mr 31,000 and 36,000. The DMSO medium proteoglycans had only the 31,000-Mr core protein, and the PMA culture medium proteoglycans had core proteins of Mr 27,000, 31,000 and 36,000. Changes in synthesis of proteoglycans induced by DMSO or PMA may have relevance for the maturation of haematopoietic cells.


Conformational differences between surface-bound and fluid-phase complement-component-C3 fragments. Epitope mapping by cDNA expression. In previous studies a subset of complement-component-C3 (C3) epitopes, C3(D), expressed in denatured and surface-bound C3 and C3 fragments, has been described. These epitopes were detected by antibodies raised against denatured C3. In the present study we used a cDNA expression strategy to localize epitopes recognized by monoclonal and polyclonal anti-C3(D) antibodies. First, DNAse I digestion of C3 cDNA was used to generate 200-300 bp fragments. These cDNA fragments were expressed as beta-galactosidase-C3 fusion proteins using the lambda gt11 vector. The fusion proteins were tested by Western-blot analysis for reactivity with monoclonal and polyclonal anti-C3 antibodies, and the location of the epitopes were determined by sequencing the cDNA fragments. Affinity-purified polyclonal anti-C3(D) antibodies specific for denatured C3 reacted strongly with the C3 fusion fragments corresponding to segments of the 40 kDa subunit of C3c (residues 1477-1510) and the C3d fragment (residues 1117-1155 and 1234-1294) of C3. Adsorption of the polyclonal antibodies with a mixture of EAC3b and EAC3bi (degradation fragments of C3 bound to sheep erythrocytes) abolished binding to fusion proteins spanning the C3d region, but not the 40 kDa fragment of C3c. No effect was seen with the corresponding soluble C3 fragments. The monoclonal anti-C3(D) antibodies (mAbs) 7D326.1 and 7D331.1, specific for EAC3b and EAC3bi, bound to a fusion protein corresponding to amino acid residues 1312-1404, whereas mAb 7D9.2, specific for EAC3d, reacted with a fusion protein spanning amino acid residues 1082-1118. mAbs 4SD11.1 and 4SD18.1, which did not bind to any physiological C3 fragment, detected a fusion protein covering residues 1477-1510. In summary, the segments of C3 represented by amino acid residues 1082-1118, 1117-1155, 1234-1294 and 1312-1404 accommodate C3(D) epitopes that are expressed by erythrocyte-bound C3 fragments, but not by the corresponding fluid-phase fragment, whereas the segments spanning residues 973-1026 and 1477-1510 contain C3(D) epitopes that are exposed exclusively in denatured C3 and therefore hidden in physiological fragments of the protein.


Phosphorylation of elongation factor 2 during Ca(2+)-mediated secretion from rat parotid acini. In this paper we report the rapid phosphorylation of a cytosolic 100 kDa protein during stimulation of secretion from dispersed aggregates of parotid acinar cells with Ca(2+)-mobilizing secretagogues (carbachol, Substance P, ATP and the Ca2+ ionophore A23187). Phosphorylation was inhibited by removal of extracellular Ca2+ but was not observed during stimulation with phorbol esters, suggesting that this protein is not a substrate for protein kinase C. Two-dimensional PAGE and immunoprecipitation with a specific antiserum indicated that this protein is elongation factor 2, whose Ca2+ calmodulin-dependent phosphorylation has been shown to inhibit protein synthesis [Nairn & Palfrey (1987) J. Biol. Chem. 262, 17299-17303]. These results suggest that phosphorylation of elongation factor 2 is the molecular mechanism for the inhibition of protein synthesis which has been previously observed in rat parotid cells during stimulation with Ca(2+)-mobilizing secretagogues.


Interaction between the calcium and cyclic AMP messenger systems in perifused rat parotid acinar cells. Possible mechanism for potentiation of amylase secretion. Potentiation of amylase secretion induced by a combination of isoproterenol and carbamylcholine was examined in perifused rat parotid acinar cells. The time course of changes in the augmented amylase secretion induced by isoproterenol plus carbamylcholine was similar to that induced by carbamylcholine alone, but not to that caused by isoproterenol. Concentration-response analysis showed that isoproterenol increased the apparent affinity for carbamylcholine to stimulate amylase secretion with the maximum effect attained by isoproterenol plus carbamylcholine being higher than that attained by isoproterenol or carbamylcholine. 8-Bromo cyclic AMP, forskolin and 3-isobutyl-1-methylxanthine mimicked the effect of isoproterenol. Calcium ionophores (A23187 and ionomycin), but not phorbol 12,13-dibutyrate, mimicked the effect of carbamylcholine. Chelation of intracellular free calcium with 1,2-bis-[2-aminophenoxyl]-ethane-N,N,N',N'-tetraacetic acid, but not that of extracellular calcium with [ethylenebis(oxyethylenenitrile)]-tetraacetic acid (EGTA), abolished the potentiation. Calmodulin antagonists inhibited amylase secretion induced by isoproterenol plus carbamylcholine or carbamylcholine alone, but not that induced by isoproterenol alone. These results suggest that the potentiation is mainly, if not completely, caused by a coordinated interaction between the cyclic AMP system and the Ca2+ system at a step distal to second messenger generation, probably via a cyclic AMP-induced increase in the sensitivity of the Ca2+ response element to calcium.


A pleiotropic response to phenobarbital-type enzyme inducers in the F344/NCr rat. Effects of chemicals of varied structure. The effects of a number of phenobarbital-type inducers on selected drug-metabolizing enzymes in male F344/NCr rats were determined by measuring specific catalytic activities and/or by measuring the levels of RNA which hybridize with specific probes for the corresponding genes. The effects on hepatic CYP2B1 were assessed by measuring the levels of CYP2B1-specific RNA and benzyloxyresorufin O-dealkylase and testosterone 16 beta-hydroxylase activities. Levels of CYP3A were monitored by measuring the rate of hydroxylation of testosterone at the 6 beta-position. Microsomal epoxide hydrolase activity was determined by measurement of cellular RNA specific for this form and by assaying the hydrolysis of benzo[a]pyrene-4,5-oxide. UDP-glucuronyltransferase activity was assayed by measuring the glucuronidation of 3-hydroxybenz[a]anthracene. Levels of glutathione S-transferase Ya/Yc were measured by quantifying total cellular RNA coding for the proteins. When male F344/NCr rats were administered various doses of phenobarbital or dichlorodiphenyltrichloroethane (DDT), strong correlations between the induction of CYP2B1 and the induction of epoxide hydrolase or UDP-glucuronyltransferase activities were observed. Treatment of rats with barbiturates, hydantoins, halogenated pesticides such as DDT or alpha-hexachlorocyclohexane, 2,4,5,2',4',5'-hexachlorobiphenyl, CYP2B1 inhibitors such as clotrimazole or clonazepam, or such structurally-diverse compounds as 2-hexanone or diallyl sulfide resulted in induction of CYP2B1-mediated enzyme activity and induction of certain other forms of cytochrome P450, microsomal epoxide hydrolase, at least one form of UDP-glucuronyltransferase, and multiple forms of glutathione S-transferase. This suggests that, as a class, compounds which induce CYP2B1 also induce a coordinate hepatic pleiotropic response which includes induction of these other phase I and phase II drug-metabolizing enzymes.


Effects of anti-allergic drugs on human neutrophil superoxide-generating NADPH oxidase. The effects of anti-allergic drugs with or without H1-receptor antagonism on the NADPH oxidase (EC 1.6.99.6) from human neutrophils in both whole-cell and fully soluble (cell-free) systems were investigated. Three anti-allergic drugs with H1-receptor antagonism, azelastine, ketotifen and oxatomide, were found to inhibit the superoxide generation of human neutrophils exposed to phorbol myristate acetate in a whole-cell system and the activation of superoxide-generating NADPH oxidase by sodium dodecyl sulfate in a cell-free system. The concentrations of these drugs required for 50% inhibition of the oxidase (IC50) were: azelastine--0.7 microM in the whole-cell system and 0.5 microM in the cell-free system; ketotifen--60 microM in the whole-cell system and 6.8 microM in the cell-free system; and oxatomide--25 microM in the whole-cell system and 9.7 microM in the cell-free system. In addition, in the cell-free system, these drugs did not change the Km values for the NADPH of the oxidase. However, these drugs did not inhibit the superoxide generation of NADPH oxidase after its activation in whole-cell and cell-free systems, suggesting that these drugs do not have superoxide-scavenger actions. Concentrations of less than 200 microM of anti-allergic drugs without H1-receptor antagonism, tranilast, repirinast and ibudilast, did not inhibit neutrophil NADPH oxidase in whole-cell and cell-free systems. The IC50 of hydrocortisone in the cell-free system was 60 microM. These results suggest that anti-allergic drugs with H1-receptor antagonism inhibit activation of the solubilized membrane-bound enzyme by sodium dodecyl sulfate in cell-free systems and that they have much stronger anti-inflammatory action than hydrocortisone.


Modulation of melphalan uptake in murine L5178Y lymphoblasts in vitro by changes in ionic environment. The alkylating agent melphalan is actively transported in mammalian cells by two amino acid transport carriers: the sodium-dependent carrier with substrate preference for alanine-serine-cysteine (system ASC), and a sodium-independent carrier with preference for leucine (system L). The effect of altering the ionic environment of murine L5178Y lymphoblasts was investigated in order to determine not only the direct effects of hydrogen and calcium ions on these transport systems, but also the indirect effects of agents or modulators known to alter intracellular calcium. Melphalan transport followed a bell-shaped distribution curve over a pH range from 3 to 9 with a pH optimum of 4.3 and 4.6 for transport by systems ASC and L, respectively. Those agents that could cause a decrease in cytosolic calcium such as the calcium channel blockers verapamil, diltiazem and nitrendipine, the calcium chelator (ethyleneglycol-bis-(beta-aminoethylether) N,N,N',N'-tetraacetic acid (EGTA) and reduction of pH were found to augment melphalan uptake, whereas conditions that would elevate intracellular calcium such as the calcium ionophore A23187, the calcium channel agonist (-) Bay K 8644, elevation of extracellular calcium and the calcium pump inhibitor trifluoperazine were all found to decrease melphalan uptake. These findings suggest that modification of ionic environment directly or indirectly by agents known to alter intracellular calcium can modulate melphalan uptake.


Highly acetylated H4 is associated with histone displacement in rat spermatids. The presence of highly acetylated histone H4 during spermatogenesis was studied to evaluate its correlation with the events of gene transcription, histone deposition, and histone displacement. We utilized an antibody raised to a pentaacetylated synthetic peptide that preferentially recognizes highly (tetra- and tri-) acetylated forms of rat testis H4. Electrophoretic separation of histones from enriched fractions of spermatogenic cells followed by detection of these forms by staining and by immunoblotting using this antibody showed that the highly acetylated forms were limited almost exclusively to spermatids beginning at step 11 of development. Immunoflurescence also revealed a striking polarity in the progression of histone from the spermatid nucleus. Highly acetylated H4 was displaced from the anterior to the caudal portion of the spermatid nucleus during steps 11 and 12, along with other histones, prior to their displacement by transition proteins. Thus, while monoacetylated and low levels of diacetylated forms of H4 were associated with stages at which histone deposition and transcription occur, the more highly acetylated forms appeared in high levels only at the stage at which histone displacement occurs.


Effect of fetal haemoglobin on the accuracy of pulse oximetry in preterm infants. Pulse oximeters are programmed with a calibration curve derived from studies done in adults. Whether fetal haemoglobin levels affect their reliability is unclear. This study reports the accuracy of pulse oximetry in 22 preterm infants (mean 31 weeks, range 25-36 weeks gestation) between 1 h and 73 days of age. Oxygen saturation obtained from a Nellcor N-200 pulse oximeter (SpO2) was compared with simultaneous arterial values (functional SaO2) measured by a Radiometer OSM3 Hemoximeter over a SpO2 range of 83-99%. Fetal haemoglobin (HbF), carboxyhaemoglobin (HbCO) and methaemoglobin (HbMet) measured by the hemoximeter ranged between 0-100%, 0-3.5% and 0-0.8% respectively. Linear regression analysis revealed a close correlation between SpO2 and functional SaO2 (SpO2 = 0.75 SaO2 + 24.43, r = 0.88, P less than 0.001) over a wide range of values for PCV, heart rate, blood pressure, PaO2, PaCO2 and pH. The mean SpO2-SaO2 difference of 1.3, (s.d. 2.5%, P less than 0.001) was unaffected by HgF, HbCO or HbMet but was increased in infants receiving inotropic support. We conclude that the Nellcor N-200 pulse oximeter gives reliable oxygen saturation measurements unaffected by the HbF level in preterm infants.


The role of insulin-sensitive phosphodiesterase in insulin action. The physiological role of the insulin-sensitive phosphodiesterase in mediating the antilipolytic actions of insulin was investigated in rat fat cells with two phosphodiesterase inhibitors, namely, 3-isobutyl-1-methylxanthine (IBMX) and griseolic acid. Insulin activates the phosphodiesterase when incubated with intact fat cells and blocks isoproterenol-induced cellular cAMP production and lipolysis, in a time- and dose-dependent manner. High concentrations of IBMX (1 mM), but not lower concentrations (0.1 mM), increased fat cell cAMP levels and lipolytic responses and overcame the antilipolytic effects of insulin; however, this treatment does not inhibit insulin-stimulated phosphodiesterase activity at earlier times (less than 30 min of incubation). These results may suggest that the level of cAMP under these conditions may be sufficient to stimulate lipolysis maximally, even though increases in cellular cAMP accumulation associated with the higher concentration of IBMX (1 mM) are partially suppressed by insulin. Cellular cAMP accumulation and the phosphodiesterase activation induced by IBMX are suppressed by the nonhydrolyzable adenosine analogue N6-phenylisopropyladenosine (PIA). These results suggest the involvement of adenosine receptors in mediating these responses. A novel phosphodiesterase inhibitor, griseolic acid, suppresses basal phosphodiesterase activity (approximately 50%), increases cellular cAMP content, and stimulates lipolysis in intact fat cells. It also partially suppresses insulin-stimulated phosphodiesterase activity (approximately 50%) and reduces the ability of insulin to decrease cellular cAMP concentration (approximately 40%) and lipolysis (approximately 65%). Because, unlike IBMX and other drugs, griseolic acid demonstrates only inhibition of phosphodiesterase activity, it may be a useful tool for studying the mechanism of insulin action in intact cells. The present work supports the concept that insulin's antilipolytic action is mediated by an activation of fat cell phosphodiesterase.


Stenting of the arterial duct: a new approach to palliation for pulmonary atresia. To assess the possibility of maintaining ductal patency in neonates with complex pulmonary atresia by percutaneous implantation of balloon expandable stents. Two duct-dependent neonates with long segment pulmonary atresia, right sided aortic arch, and left sided arterial duct. Stents with final diameter of 3.5 or 4 mm and initial length of 7 or 15 mm were successfully positioned in the arterial duct. Two stents were required in one child and four in the other in order to stent the entire length of the duct. After the procedures the ducts remained widely patent and arterial oxygen saturations remained above 80%. Complications of the procedures included perforation of a peripheral pulmonary artery and cardiac perforation, both caused by guide wire manipulation. Both babies died suddenly, one at five weeks, and the other at nine days after successful stenting of the duct. Both ducts were patent at necropsy; the exact cause of one death was not clearly defined, but the second seemed to be caused by pneumococcal septicaemia. Stenting of the arterial duct is technically feasible. It provides adequate palliation for neonates with pulmonary atresia at least in the short term and it seems to result in balanced, central perfusion of both pulmonary arteries. This preliminary report suggests that this previously untried technique may prove to be a promising and attractive alternative to neonatal aortopulmonary shunt operation.


Eccrine syringofibroadenoma: a case report with analysis of cytokeratin expression. A 56-year-old man presented with a 30-year history of a slowly enlarging lesion on the sole of his right foot. A biopsy showed an anastomosing network of small cuboidal cells with the formation of occasional sweat ductal lumina and a marked fibrovascular stroma. The histological findings were interpreted as consistent with the diagnosis of an eccrine syringofibroadenoma. Using immunohistochemistry all the tumour cells were positively stained by the pan-cytokeratin antibody Lu-5 and an antibody to the cytokeratins 1/5/10/11. In addition the luminal ductal cells expressed cytokeratin 19 and CEA. Tumour cells were negative for cytokeratins 1, 7, 8, 13 and 18 and did not express vimentin and GCDFP-15. The results indicate that the eccrine syringofibroadenoma is differentiated towards the dermal eccrine duct.


Palliation of bone metastases in prostate cancer. Hemibody irradiation or strontium-89? The palliation of bone pain is a common clinical problem once metastatic prostate cancer has escaped from hormonal control. This retrospective study compares the results of treatment using hemibody irradiation (HBI) at the Royal Marsden Hospital (27 cases) with isotope therapy using the bone-seeking isotope strontium-89 (89Sr) at Southampton General Hospital (51 cases). Prior to analysis patients were matched for potential prognostic factors (performance status, bone scan extent of disease, age, histology and duration of hormone response) to minimize the effect of treatment selection bias. Pain control assessed at 3 months was similar for HBI and matched 89Sr cases, with 63% and 52% respectively showing some benefit. Median survival was similar for these groups at 20 and 21 weeks respectively. The unmatched 89Sr group, which had more favourable prognostic factors, had a better outcome with 96% showing improvement in pain and with a median survival of 59 weeks. Subsequent univariate analysis demonstrated that performance status and extent of disease on bone scan were of overriding importance in determining outcome. Transfusion requirements were higher for the HBI group than for the matched 89Sr group (50% and 25% respectively) but other bone marrow toxicity was similar. Despite routine anti-emetic therapy 37% of patients treated with HBI had some nausea or vomiting. Although expensive, 89Sr appears as effective a treatment option as HBI. Response is most likely with either approach when patients have a good performance status and a limited extent of disease.


The role of radiotherapy in carcinoma of the thoracic oesophagus: an audit of the Mount Vernon experience 1980-1989. All 244 patients with carcinoma of the thoracic oesophagus registered at the Mount Vernon Centre for Cancer Treatment during the decade from 1 January 1980 to 31 December 1989 have been audited. We have made a detailed analysis of 110 (45%) with localized disease considered unsuitable for surgery, who completed treatment solely by radiotherapy. The median survival of this group of patients was 8.2 months (range 0.2-54 months). Dysphagia was improved by radiotherapy in 77.3% of cases, the median duration of relief was 24 weeks (range 0-208 weeks) and was maintained until death in 40%. Life table analysis showed that radical compared with less than radical regimens of radiotherapy gave significantly superior relief of dysphagia. This result is unlikely to be due to case selection.


Metastasis to the penis from malignant melanoma: case report and review of the literature. A case of metastatic malignant melanoma to the shaft of the penis is described and the literature reviewed to collate the incidence of primary sites which metastasize to the penis. Less than 260 cases of metastasis to the penis have been reported. Of these, 76% are from genitourinary primary sites and 17% are from gastrointestinal primary sites but only one case of metastatic melanoma to the penis has been previously reported. The described case presented with painful priapism while receiving combination chemotherapy for metastatic disease. A CT scan demonstrated a deposit in the left corpora cavernosa and needle aspiration cytology of a plaque attached to the shaft confirmed malignant melanoma cells. Palliation of the painful priapism was achieved by treatment with radiotherapy using large doses per fraction. Retrograde venous or lymphatic spread may have been the cause of a metastasis at this site. Prognosis is very poor.


Maternal serum screening for Down's syndrome: the effect of routine ultrasound scan determination of gestational age and adjustment for maternal weight. To investigate the effect of using a routine ultrasound estimate of gestational age and maternal weight adjustment on maternal serum alpha-fetoprotein (AFP), unconjugated oestriol (uE3) and human chorionic gonadotrophin (hCG) levels in antenatal screening for Down's syndrome. Women with a singleton pregnancy without Down's syndrome were screened using the three serum markers and an estimate of gestational age based on 'dates' (time since first day of the last menstrual period) and one based on an ultrasound scan examination was recorded together with maternal weight. Women attending the Homerton Hospital, Hackney, for their antenatal care between February 1989 and January 1990. 2113 women with a singleton pregnancy without Down's syndrome. The use of ultrasound to estimate gestational age (usually based on the biparietal diameter of the fetal skull) led to a significant reduction in the variance of each marker at a given week of pregnancy. The level of each marker was negatively associated with maternal weight, so that adjustment for weight also led to a reduction in variance. These data on gestational age and maternal weight, taken together with published data on pregnancies associated with Down's syndrome, indicate that the routine use of ultrasound to estimate gestational age will increase the detection rate from 58% to 67% while maintaining the false-positive rate at 5%, or reduce the false-positive rate from 5.7% to 3.1% while maintaining the detection rate at 60%. Routine maternal weight adjustment for the serum marker levels was much less useful, increasing the detection rate by about 0.5% for a given false-positive rate, or reducing the false-positive rate about 0.1% for a given detection rate. An ultrasound gestational age estimate available at the time of Down's syndrome screening confers a substantial advantage to screening performance with a further small benefit resulting from maternal weight adjustment, which is worth adopting if it can be done without difficulty or extra cost.


Changes in the methylation pattern of the TCR zeta-chain gene correlate with its expression in T cells and developing thymocytes. The tight regulation of T-cell gene expression during thymic ontogeny is essential to the development of a normal immune system. One set of developmentally regulated genes encodes the multicomponent T-cell antigen receptor (TCR). The zeta chain, a component of the TCR, has been shown to play important roles in signal transduction from antigen binding to T-cell activation and in transport of the TCR complex to the cell surface. In this study, we examine the regulation of zeta gene expression in murine T-cell hybridomas. In these cells, zeta expression is correlated with complex, but predictable, changes in the pattern of cytosine methylation of its gene. Some of these structural changes are identical to those observed in murine fetal thymocytes and correlate with the rapid alteration of zeta message seen in the thymus between days 15 and 18 of gestation.


Multiple binding sites for tetrahedral oxyanion inhibitors of bovine spleen purple acid phosphatase. The theory of multiple inhibition kinetics has been extended to enzymes for which one inhibitor is noncompetitive and the other exhibits mixed inhibition. Plots of reciprocal velocity versus the concentration of either inhibitor at various fixed concentrations of the second inhibitor are predicted to give parallel lines if binding of the inhibitors is mutually exclusive and intersecting lines if the inhibitors interact at different sites on the enzyme. Application of this analysis to the purple acid phosphatase from bovine spleen in the presence of molybdate (a noncompetitive inhibitor) and phosphate (which exhibits mixed inhibition) results in parallel lines in the reciprocal velocity plots, indicating that phosphate and molybdate compete for a common site; since molybdate is a noncompetitive inhibitor, this site is inferred to be distinct from the site at which substrate binds and is hydrolyzed. Extension of these ideas suggests that phosphate ester substrates should be capable of binding to the molybdate-binding site as well as to the active site, and evidence for substrate inhibition at high substrate concentrations has been obtained. The implications of these findings for interpretation of previous spectroscopic studies of purple acid phosphatase complexes with tetrahedral oxyanions are discussed.


DAPI (4',6-diamidino-2-phenylindole) binds differently to DNA and RNA: minor-groove binding at AT sites and intercalation at AU sites. The interaction of DAPI and propidium with RNA (polyA.polyU) and corresponding DNA (polydA.polydT) sequences has been compared by spectroscopic, kinetic, viscometric, Tm, and molecular modeling methods. Spectral changes of propidium are similar on binding to the AT and AU sequences but are significantly different for binding of DAPI. Spectral changes for DAPI with the DNA sequence are consistent with the expected groove-binding mode. All spectral changes for complexes of propidium with RNA and DNA and for DAPI with RNA, however, are consistent with an intercalation binding mode. When complexed with RNA, for example, DAPI aromatic protons signals shift significantly upfield, and the DAPI UV-visible spectrum shows significantly larger changes than when complexed with DNA. Slopes of log kd (dissociation rate constants) versus-log [Na+] plots are similar for complexes of propidium with RNA and DNA and for the DAPI-RNA complex and are in the range expected for an intercalation complex. The slope for the DAPI-DNA complex, however, is much larger and is in the range expected for a groove-binding complex. Association kinetics results also support an intercalation binding mode for the DAPI-RNA complex. The viscosity of polyA.polyU solutions increases significantly on addition of both propidium and DAPI, again in agreement with an intercalation binding mode for both molecules with RNA. Molecular modeling studies completely support the experimental findings and indicate that DAPI forms a very favorable intercalation complex with RNA. DAPI also forms a very stable complex in the minor groove of AT sequences of DNA, but the stabilizing interactions are considerably reduced in the wide, shallow minor groove of RNA. Modeling studies,thus,indicate that DAPI interaction energetics are more favorable for minor-groove binding in AT sequences but are more favorable for interaction in RNA.


Folate binding protein from kidney brush border membranes contains components characteristic of a glycoinositol phospholipid anchor. A number of cell surface proteins have been shown to be anchored to the plasma membrane by a covalently attached glycoinositol phospholipid (GPL) in amide linkage to the C-terminus of the mature protein. We applied several criteria to establish that folate binding protein (FBP) in brush border membranes of rat kidney contains a GPL anchor. Brush border membranes were isolated and labeled with [3H]folate, and the complex of FBP and [3H]folate was shown to be released to the supernatant by incubation with purified bacterial phosphatidylinositol-specific phospholipase C (PIPLC) but not by incubation with a purified bacterial phosphatidylcholine-specific phospholipase C. The FBP-[3H]folate complex both in crude extracts and after FBP purification by ligand-directed affinity chromatography interacted with Triton X-114 micelles, and prior incubation with PIPLC prevented this detergent interaction. Individual residues characteristic of GPL anchors were found to be covalently associated with FBP following polyacrylamide gel electrophoresis in sodium dodecyl sulfate. These included glucosamine and ethanolamine, which were radiolabeled by reductive methylation and identified by chromatography on an amino acid analyzer, and inositol phosphate, which was inferred by Western blotting with an anti-CRD antisera. This antisera gave positive immunostaining only after FBP had been cleaved by PIPLC, a reliable diagnostic of a GPL anchor. The relationship between GPL-anchored FBP in biological membranes and soluble FBP in biological fluids also is discussed.


Ca2+/calmodulin-dependent NO synthase type I: a biopteroflavoprotein with Ca2+/calmodulin-independent diaphorase and reductase activities. NO synthase (NOS; EC 1.14.23) catalyzes the conversion of L-arginine into L-citrulline and a guanylyl cyclase-activating factor (GAF) that is chemically identical with nitric oxide or a nitric oxide-releasing compound (NO). Similar to the other isozymes of NOS that have been characterized to date, the soluble and Ca2+/calmodulin-regulated type I from rat cerebellum (homodimer of 160-kDa subunits) is dependent on NADPH for catalytic activity. The enzyme also possesses NADPH diaphorase activity in the presence of the electron acceptor nitroblue tetrazolium (NBT). We investigated the requirements of NOS and its content of the proposed additional cofactors tetrahydrobiopterin (H4biopterin) and flavins, further characterized the NADPH diaphorase activity, and quantified the NADPH binding site(s). Purified NOS type I Ca2+/calmodulin-independently bound the [32P]2',3'-dialdehyde analogue of NADPH (dNADPH), which, at near Km concentrations during 3-min incubations was utilized as a substrate and at higher concentrations or after prolonged incubations and cross-linking inhibited NOS activity. The NADPH diaphorase activity was Ca2+/calmodulin-independent, required higher NADPH concentrations than NOS activity, and was affected by dNADPH to a lesser degree. Divalent cations interfered with the diaphorase assay. Per dimer, native NOS contained about 1 mol each of H4biopterin, FAD, and FMN, classifying it as a biopteroflavoprotein, and incorporated 1 mol of dNADPH. No dihydrobiopterin (H2biopterin), biopterin, or riboflavin was detected. These findings suggest that NOS may share cofactors between two identical subunits via high-affinity binding sites.(ABSTRACT TRUNCATED AT 250 WORDS)


Structural and functional characterization of the phosphorylated adipocyte lipid-binding protein (pp15). A substrate for the insulin receptor kinase in 3T3-L1 adipocytes has previously been identified as the adipocyte lipid-binding protein (ALBP, also known as aP2 or p15). We have characterized the effect of tyrosyl phosphorylation on ALBP structure and ligand-binding properties. Phosphorylated ALBP (phospho-ALBP) was isolated by a combination of gel filtration, anion exchange chromatography, and immunoaffinity chromatography on anti-phosphotyrosine agarose. Circular dichroic spectroscopy indicated that the phosphoprotein was similar in structure to native ALBP. Phospho-ALBP exhibited a slight decrease in calculated alpha-helical content which was compensated for by an increase in beta-sheet structure. The wavelength yielding maximum tryptophan fluorescence was unaltered by phosphorylation (334 +/- 1 nm). However, the concentration of guanidine HCl yielding 50% denaturation was 1.43 M for ALBP and 0.92 M for phospho-ALBP. The delta Goapp was 3.87 and 3.25 kcal mol-1 for ALBP and phospho-ALBP, respectively, suggesting that phosphorylation destabilized the protein. To assess the binding characteristics of the phosphoprotein, a long-chain fatty acid affinity column was synthesized to which native ALBP specifically bound. In contrast, phospho-ALBP showed little or no affinity for the column. Furthermore, phosphorylation virtually abolished binding of the fluorescent fatty acid analogue 12-(9-anthroyloxy)oleic acid. Fatty acid binding activity was recovered (approximately 60%) upon dephosphorylation with protein tyrosine phosphatase. The structural studies, coupled with the crystal structure of the apoprotein, indicate that the dramatic reduction in binding affinity is likely a result of steric hindrance in the binding cavity or of electrostatic interactions of the phosphoryl group with the fatty acid.(ABSTRACT TRUNCATED AT 250 WORDS)


Vitronectin diversity in evolution but uniformity in ligand binding and size of the core polypeptide. We isolated vitronectins from the plasma or sera of 14 animal species including mouse and rat by heparin affinity chromatography. They cross-reacted with anti-vitronectin antibody and their amino terminal sequences showed strong homology. They also promoted spreading of BHK cells and were bound to heparin and collagen in the same way. Therefore, these properties appear to be essential for vitronectin function. However, the apparent molecular weights of these vitronectins varied considerable from 59 to 78 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In addition, the number of bands also varied from 1 to 3. To search for the uniformity of vitronectin polypeptide, vitronectins were deglycosylated and examined by Ferguson plot analysis. The size of the polypeptide portion of vitronectins was estimated to range from 40 to 57 kDa which was 19-26 kDa smaller than original values. Supposing a possible cleavage site at 5-13 kDa far from the carboxyl terminus, all vitronectin polypeptides were speculated to be synthesized de novo in the size range of 50-57 kDa. Proteins reacting with anti-vitronectin antibody were also detected on the immunoblot of 13 more species including Drosophila and Physarum. Almost all of these vitronectin-like proteins showed marked species-specific variations in their apparent molecular weights from 51 to 96 kDa in SDS-PAGE.


Separation and characterization of isoforms of DT-diaphorase from rat liver cytosol. Rats were treated with 3-methylcholanthrene (MC) and DT-diaphorase from liver was partially purified on an azodicoumarol-Sepharose 6B column and applied to an FPLC-chromatofocusing column in order to resolve isoforms. Six peaks showing significant DT-diaphorase activity were eluted from this column with a pH gradient between 7.30 to 4.80. The amino acid compositions of the two major peaks (II and VIb) were found to be nearly identical, suggesting existence of isoforms rather than isozymes of DT-diaphorase. The isoforms of DT-diaphorase showed broad substrate specificities towards four different quinones (menadione, vitamin K-1, benzo(a)pyrene 3,6-quinone and cyclized-dopamine ortho-quinone), although quantitative differences in the specific activities were also found. All isoforms are glycoproteins but contain different carbohydrates. Thus isoform II reacts with biotinylated lectins which are specific for N-acetylgalactosamine, mannose, fucose and galactosyl(beta-1,3)N-acetylgalactosamine, while isoform VIb reacts only with biotinylated lectins specific for mannose and N-acetylgalactosamine. Separation of DT-diaphorase isoforms from control rat liver cytosol using FPLC-chromatofocusing revealed that the induction of the isoforms is not uniform, since isform II was not found and the major isoform was composed of three peaks, whereas the major isoform of DT-diaphorase from liver cytosol of rats treated with 3-methylcholanthrene was composed of only two peaks.


Cyclic AMP produces desensitization of prostacyclin and adenosine A2 receptors in hybrid cell lines but does not affect Gs function. Prostacyclin and adenosine A2 receptors stimulate adenylate cyclase activity in the related somatic hybrid cell lines NG108-15 and NCB20. The role of cAMP in the desensitization of these receptors has been examined. Pretreatment for 17 h with forskolin or 8-bromo-cAMP had the same effect in both cell lines. There was no change in the response to sodium fluoride or forskolin, suggesting that the function of Gs and adenylate cyclase were unaffected by increased levels of cAMP. Receptor responses were affected however; the maximum response to N-ethylcarboxamidoadenosine (an A2 receptor agonist) was reduced by 30-40%, there was a small but consistent shift to the right of the dose-response curve for iloprost (a stable analogue of prostacyclin) and [3H]iloprost binding studies revealed a loss of prostacyclin receptors. However, the loss of receptor responsiveness was much smaller than that which occurs following pretreatment with prostacyclin or adenosine A2 receptor agonists (Keen et al. (1989) Biochem. Pharmacol. 38, 3827-3833; Kelly et al. (1990) Br. J. Pharmacol. 99, 309-316) suggesting that cAMP may not play a major role in agonist mediated desensitization.


Immunohistochemical detection of porcine rotavirus using immunogold silver staining (IGSS). Immunogold silver staining (IGSS) was applied for the detection of porcine group A rotavirus in formalin-fixed paraffin-embedded tissue sections of small intestine. Prior to the application of IGSS, the reactivity of protein A-gold as a marker was tested with group-specific antiserum in immunogold electron microscopy. Immune aggregates were intensely and specifically labeled with the gold complex. Application of IGSS to tissue sections resulted in specific dark staining of villous enterocytes infected by group A rotavirus. This method also proved effective for the detection of rotaviral antigen in infected cultured cells. The IGSS method may be suitable for routine diagnostic detection of rotaviral infections and may have application for detection of other viral pathogens of veterinary importance.


Phenotypic and clinical heterogeneity of CD56-positive acute nonlymphoblastic leukemia. The precise phenotype and clinical course are described of a subgroup of acute nonlymphoblastic leukemias (ANLL) expressing the NK-cell differentiation antigen CD56. As previously reported, CD56+ leukemias occurred in a frequency of about 20% of ANLL cases showing clinical and immunophenotypical heterogeneity. Carrying various myelomonocytic markers, all cases were diagnosed to be of nonlymphoid origin. Positive or negative expression of CD34 allowed us to distinguish two major subtypes of CD56+ leukemias representing immature and more differentiated cells carrying further differentiation antigens (CD14 and/or CD15) of the myelomonocytic lineages. These phenotypes correlated with the M0, M2, M4, and M5 leukemias of the FAB classification.


Protein structural effects in gas phase ion/molecule reactions with diethylamine. The relationship between gas-phase protein structure and ion/molecule reactivity is explored in comparisons between native and disulfide-reduced aprotinin, lysozyme, and albumin. Reactions are performed in the atmospheric-pressure inlet to a quadrupole mass spectrometer employing a novel capillary interface-reactor. In reactions with equal concentrations of diethylamine, multiply protonated molecules generated by electrospray ionization (ESI) of 'native' proteins shifted to lower charge states than did multiply protonated molecules from ESI of the disulfide-reduced counterparts, suggesting that the disulfide-reduced protein ions are less reactive than native protein ions of the same charge state. Differences in reactivity may arise from protonation of different amino acid residues and/or differences in the proximities of charge sites in the two molecules. These results suggest that the reactivity of multiply charged proteins can be significantly affected by their gas-phase structure.


Integrin expression by human articular chondrocytes. Expression of integrins, a family of cell adhesion proteins, by human articular cartilage chondrocytes in vivo has been studied by immunohistological techniques using cryostat sections and a panel of anti-integrin monoclonal antibodies. Chondrocytes strongly expressed beta 1 and alpha 5 integrin subunits, known to form the classical fibronectin receptor, VLA-5, strongly. alpha 1 was expressed more weakly and variably; alpha 3 was expressed by occasional cells only. Chondrocytes did not express beta 2, beta 3 or other integrin alpha subunits tested.


Constitutional translocation (8;13) in a patient with non-Hodgkin's lymphoma. We report a case of non-Hodgkin's lymphoma in a 16-year-old male, whose peripheral white blood cells have a t(8;13)(q24;q14). There are no previous reports that describe this association. Although the tumor cells were not studied, we discuss the possible link between this finding and the development of the malignant lymphoma.


The use of fluorescent nuclear dyes for the study of blood vessel structure and function: novel applications of existing techniques. We have used nuclear fluorescent dyes to develop a technique for the study of vascular structure and function. Nuclear stained blood vessels, viewed with the appropriate filter sets, can be studied in great detail. Only the nuclei of the cells which form the walls are visible and so their positions relative to one another as well as their viability can be quickly assessed. The dyes are not toxic, therefore when the vessel contracts or relaxes, the changes in position of the nuclei can be monitored. In this paper we describe two original applications of fluorescent nuclear dyes in vascular research.


Marrow fibrosis associated with a Philadelphia chromosome. Three patients had marked marrow fibrosis and an apparent Philadelphia (Ph) chromosome. Hematologic, cytogenetic, and molecular studies demonstrated the heterogeneity of such cases, including the first example of clinically typical myelofibrosis (MF) associated with a bcr gene rearrangement characteristic of chronic myelogenous leukemia (CML).


Potentiation of transmembrane signaling by cross-linking of antibodies against the beta chain of the T cell antigen receptor of JURKAT T cells. Three monoclonal antibodies (mAb) 2D1, 3B9, and 3B12 were produced by immunizing BALB/c mice with JURKAT cells. These mAb induce comodulation of the TCR/CD3 complex expressed on JURKAT cells, but do not react with the CD3- JURKAT variant, J.RT3.T3.1. Immunoprecipitation studies with detergent-solubilized JURKAT cell lystes indicate that these mAb react with proteins having characteristics of the TCR molecules. Their low reactivity with peripheral blood mononuclear cells (PBMC) and lack of reactivity with other CD3+ T cell lines suggest that they may be anti-idiotypic mAb. Results from binding inhibition assays, reactivity with PBMC, and generation of transmembrane signals suggest that these three anti-TCR mAb recognized different epitopes on the TCR beta chain of JURKAT cells. Although the three mAb are capable of inducing the production of inositol phosphates and cytosolic free Ca2+ increase in JURKAT cells, their stimulatory capacities vary and are lower than that observed by anti-CD3 antibody (OKT3) stimulation. However, crosslinking these mAb with rabbit antimouse immunoglobulins potentiates the stimulatory response to comparable levels induced by OKT3. These mAb could be useful as tools to study V beta 8+ T cells in relation to antigen-specific activation.


Chromaffin cell transplants for alleviation of chronic pain. Treatment of intractable pain with parenteral, subarachnoid, or epidural narcotics is often unsatisfactory due to tolerance and other systemic complications that accompany increasing dosages of these drugs. Other disadvantages include the potential infections with implantable pumps and the inconvenience of repeated narcotic administration. During the past several years, studies at the author's laboratory indicated that transplantation of adrenal medullary tissue or isolated chromaffin cells into the spinal subarachnoid space can significantly reduce pain in several rodent models without resulting in development of tolerance. Adrenal medullary chromaffin cells were selected because they produce high levels of both opioid peptides and catecholamines, agents that independently, and possibly synergistically, reduce pain when injected locally into the spinal subarachnoid space. The adrenal medullary transplants survive for prolonged periods, and continue to produce high levels of both catecholamines and met-enkephalin. These transplants reduce pain in two rodent chronic pain models, an arthritis model and a peripheral neuropathy model, both of which closely resemble human chronic pain syndromes. The success of the animal studies has led to initiation of human clinical trials in patients with chronic cancer pain; results are promising.


CD7 augments T cell proliferation via the interleukin-2 autocrine pathway. The role of CD7, a T cell differentiation antigen, in T cell function is not known at present; this study evaluates the effect of anti-CD7 mAb in PMBC cultures activated with suboptimal concentrations of lectins, antigens, and anti-CD3 mAb. We found that the inclusion of anti-CD7 resulted in increased IL-2 production and IL-2R-alpha expression in these cultures. H-7, a protein kinase C (PKC) inhibitor, and genistein, a protein tyrosine kinase (PTK) inhibitor, significantly suppressed the proliferation of T cells in comitogenic assays. This suggested that the comitogenic effect mediated by CD7 molecule involved both the PKC and the PTK pathways of T cell activation. These drugs appeared to affect the CD7-mediated effects by inhibiting the IL-2 autocrine pathway, especially the up-regulation of IL-2R-alpha since inhibition was not relieved with exogenous rIL-2. Taken together, our results suggest that CD7 augments T cell function by up-regulating IL-2R-alpha expression and IL-2 production via multiple pathways of protein phosphorylation.


Synthetic peptides representing sequence 39 to 59 of rat V beta 8 TCR fail to elicit regulatory T cells reactive with V beta 8 TCR on rat encephalitogenic T cells. Subpathogenic doses of syngeneic autoreactive T cells protect experimental animals against associated autoimmune disease. Preferential use of the TCR of encephalitogenic T cells suggests that this molecule serves as the target for immunoregulation in experimental autoimmune encephalomyelitis (EAE). Whether peptides derived from the V beta 8 of the rat TCR elicit regulatory T cells and produce the same vaccinating effect against EAE as do whole T cells remains unknown. Here we show that immunization of Lewis rats with V beta 8(39-59), a peptide representing residues 39 to 59 of the rat V beta 8 TCR, does not induce the production of regulatory T cells reactive to the intact TCR V beta 8 containing this sequence. Moreover, animals that had recovered from both actively induced EAE and transferred EAE did not generate regulatory T cells that recognized the V beta 8(39-59) peptide. Further, transfusion of large doses of peptide-specific T cells did not protect the animals from EAE. Our results suggest that the V beta 8(39-59) peptide may comprise so-called cryptic epitopes, which function as immunogens only when dissociated from large protein complexes.


An antigen expressed by avian neuronal cells is also expressed by activated T lymphocytes. A monoclonal antibody, anti-BEN, initially characterized by its reactivity with an epitope present on the surface of avian bursa epithelial cells and neurons, also reacts with membrane molecules on some hemopoietic cells. In this study we examine BEN expression on lymphoid cells in thymus, spleen, and blood. We demonstrate that BEN is an activation antigen on mature T lymphocytes. It is not expressed on peripheral blood or splenic lymphocytes, but following mitogenic or allogeneic stimulation of blood lymphocytes it appears rapidly on a T cell subpopulation in parallel with the appearance of IL-2 receptors. BEN is also expressed on III-C5 cells, an avian IL-2-dependent permanent T cell line, and on immature CD4+CD8+ thymocytes. BEN is not expressed by resting or actively proliferating B cells. Biochemical analyses of the BEN protein on T lymphoblasts shows that the molecule is similar in size to the BEN molecules on bursa epithelial cells and on neurons. The physicochemical properties of the BEN protein and its tissue distribution differs from other known avian and mammalian T cell activation markers, differentiation antigens, and integrins. Thus BEN is a novel marker of activated T cells in birds.


Influence of feeding paradigm in rats on temporal pattern of: II. Brain serotoninergic and catecholaminergic systems. The relationship between the feeding paradigm (single diet versus food selection) and central idoleamines and catecholamines was studied. Temporal patterns of the brain parameters in response to presentation of a single diet of fixed composition (20% casein) or a choice between two isocaloric diets (0% and 60% casein) for 2 weeks under 8-h feeding cycles during the dark phase were measured in adult Sprague-Dawley rats. Groups of animals were then killed at the beginning and at 2-h intervals throughout the feeding period. The distribution and the temporal pattern of variation of the serotoninergic and the catecholaminergic parameters studied were significantly affected by the diet paradigms used. A different neurochemical equilibrium was observed before food intake and was characterized by a central serotoninergic predominance in subjects having a dietary selection experience but a central catecholaminergic predominance in animals adapted to a single diet. Hypothalamic and extrahypothalamic serotoninergic and catecholaminergic systems were found to intervene in an interdependent way, sometimes antagonistic according to the feeding paradigm and the related temporal changes in energy intake and macronutrient selection. These results suggest that central serotoninergic and catecholaminergic systems are influenced by the diet paradigm and display characteristic patterns of temporal variations during the feeding cycle. The feeding paradigm, per se, should then be considered as a potential synchronizer of central biological rhythms of monoamines, which in turn may affect food intake and appetite for macronutrients.


Induction of ventricular fibrillation versus monomorphic ventricular tachycardia during programmed stimulation. Role of premature beat conduction delay. Premature stimuli can cause ventricular fibrillation (VF) during electrophysiological testing. The electrophysiological correlations associated with the onset of VF were evaluated in 40 patients who had this rhythm induced during programmed ventricular stimulation. These parameters were compared with those observed in 51 patients who had inducible sustained monomorphic ventricular tachycardia (VT) and 45 patients who had no inducible sustained ventricular tachyarrhythmias. Shortest premature coupling intervals for S2, S3, and S4 at induction of tachycardia or before achieving refractoriness, corresponding conduction latencies (defined as the time from the premature stimulus to the upstroke of the depolarization wave front recorded 35 mm away from the stimulation site), and ventricular activation times (defined as the time from the premature stimulus to the end of the depolarization wave) were compared. The mean coupling intervals were longest in the inducible VT patients: 300 +/- 30, 254 +/- 57, and 228 +/- 32 msec for S2, S3, and S4, respectively. In the inducible VF group, the coupling intervals were 260 +/- 37, 208 +/- 20, and 213 +/- 30 msec. In the group with no inducible VT or VF, these coupling intervals were 251 +/- 24 (p less than 0.01 versus inducible VT group), 209 +/- 27 (p less than 0.001 versus inducible VT group), and 194 +/- 21 msec (p less than 0.05 versus inducible VT and VF groups). The coupling interval of the last premature extrastimulus was above 200 msec in 70% of the patients in whom VF was induced. The largest increases in latency and activation times were recorded in patients in whom VF was induced. The cumulative increase in latency, defined as increased conduction time from baseline, summed for all the premature stimuli was also the greatest at initiation of VF. In contrast, the smallest increases in these parameters were noted in the patients with no inducible VT or VF. Measurements of total activation time yielded similar results as those recorded for latencies. The most important parameters distinguishing the VT patient population from the other two groups were the low ejection fractions and the longer coupling intervals at which VT was induced, whereas in the VF group, the most important discriminating factor was cumulative activation time. Sixty-three percent of the inducible VF patients presented with abnormal hearts (myocardial infarction or cardiomyopathy), whereas 88% of the inducible VT patients had abnormal hearts. In contrast, only 25% of the patients in whom no arrhythmia was induced presented with abnormal hearts. Mean ejection fraction was 32 +/- 15% for the inducible VT group, 45 +/- 13%* for the inducible VF group, and 51 +/- 17%* for patients with no inducible VT/VF (*p less than 0.001 versus VT). The results suggest that 1) initiation of ventricular tachycardia during programmed ventricular stimulation occurs with minimal conduction latency; 2) because of the large overlap in coupling intervals where VF or VT were induced, a single coupling interval cannot be recommended to adequately separate these groups; and 3) induction of VF was preceded by increased latency and prolongation of the local activation time. These parameters should not be allowed to prolong if VF is to be avoided during programmed stimulation. In addition, 4) the initiation of VF during electrophysiological studies is often associated with the presence of structural heart disease; such structural disease may promote conduction latency and the development of VF.


Imaging of vascular injury with 99mTc-labeled monoclonal antiplatelet antibody S12. Preliminary experience in human percutaneous transluminal angioplasty. To evaluate the in vivo safety, biodistribution, and diagnostic accuracy of a monoclonal Fab' antibody (S12) that is specific for the platelet membrane glycoprotein (GMP-140) expressed during platelet activation at vascular injury sites, 11 peripheral percutaneous transluminal angioplasty (PTA) patients (age, 61 +/- 8 years) with severe vascular disease had serial 99mTcS12 radionuclide imaging at 5 and 90 minutes, 4-6 hours, and 20-24 hours after a total of 23 angiographically successful PTA procedures. No acute allergic reactions or hematologic toxicity occurred. The average PTA percent angiographic diameter stenosis (DS) at all 23 sites decreased from 85 +/- 12% to 12 +/- 11%, with a mean before-to-after-PTA change of 73 +/- 14% (p less than 0.01). The mean radionuclide image-derived ratio of 99mTc S12 activity in PTA versus contralateral non-PTA arterial segments for all angioplasty sites was 1.6 +/- 0.5. Vascular 99mTc S12 antibody activity was qualitatively evident in the majority (78%) of PTA sites at 4-6 hours after injection. 99mTc S12 target-to-background (muscle) ratio equaled 2.3 +/- 0.6 at PTA sites. Nine PTA sites (39%) had residual 99mTc S12 activity at 24 hours after injection (mean PTA site-to-contralateral artery ratio, 1.5 +/- 0.4). The mean vascular 99mTc S12 activity ratios in 10 procedurally complicated (defined as extensive dilation [greater than 2 cm] or grade I or greater arterial dissection) and 13 uncomplicated PTA segments were 1.9 +/- 0.5 versus 1.2 +/- 0.1, respectively (p less than 0.01). The associated before-to-after-PTA angiographic improvement was significantly less in procedurally complicated PTA sites (66 +/- 12% versus 80 +/- 12% DS; p less than 0.01). 99mTc S12 activity is significantly increased at angiographically patent PTA sites that are procedurally complicated and are associated with less significant before-to-after-PTA angiographic improvement. 99mTc S12 monoclonal Fab' antibody imaging permits noninvasive identification of local vascular platelet activation resulting from angioplasty balloon injury in humans.


Repetitive electrical activity of the muscle membrane induced in chloride-free medium. 1. Superficial fibres of frog skeletal muscle were electrically stimulated in Ringer solution where the chloride content had been replaced by various weakly permeant anions. Changes of the membrane potential were recorded at three different time scales. The complex response was initiated by a volley of fast repetitive action potentials (10-20 ms cycle length) superimposed on the ascending phase of a transient depolarization to -35 mV. The transient depolarization was followed by a membrane potential oscillation (0.3-0.6 s cycle length). The parameters of the volley and membrane potential oscillation were not greatly affected by the substituent anion. 2. The transient depolarization was fully abolished by tetrodotoxin (3 mumol/L), but left unaffected by nifedipine (5 mumol/L), or by the replacement of extracellular Ca for Ni or Co. Tetraethylammonium (20-40 mmol/L) increased the duration and amplitude of the transient depolarization. The shape of transient depolarization was uniform in a given fibre, in spite of its marked variability under different experimental conditions. 3. Single outward current pulses applied in chloride-free solution containing tetraethylammonium (20-40 mmol/L) evoked prolonged depolarizations to positive membrane potentials accompanied by increases in the specific membrane conductance. This slow response, which was also frequently observed in the absence of TEA, was mediated by Ca ions, as it was insensitive to tetrodotoxin (3 mumol/L) but abolished by nifedipine (10 mumol/L). 4. Two populations of the muscle fibres were observed during the slow response. Some fibres repolarized completely, while others failed to produce complete repolarization but formed a plateau between -20 and -30 mV, lasting for several minutes. When the external K concentration was abruptly increased to 5 or 10 mmol/L during the plateau of the slow response, repolarization and increase in membrane conductance were observed. 5. In muscle fibres, having osmotically disrupted T-system, the duration of transient depolarization was in the range of minutes, in contrast to the range of seconds observed in intact fibres. The volley was preserved in glycerol treated fibres, however, the baseline of discharges was close to the resting potential and the rate of depolarization of the baseline was significantly less in glycerol treated than in intact fibres. 6. These results are consistent with the existence of a second stable membrane potential level in skeletal muscle between -40 and -30 mV. The depolarization and repolarization during the membrane potential oscillation and transient depolarization can be regarded as a partial or full transition, respectively, between these two stable membrane potential levels, possibly due to the conductance changes of the inward rectifier K channels.(ABSTRACT TRUNCATED AT 400 WORDS)


Role of impairment of blood supply of the femoral head in the pathogenesis of idiopathic osteonecrosis. To investigate the role of blood supply in the pathogenesis of idiopathic osteonecrosis of the femoral head, superselective angiography of the medial circumflex artery was performed. Sixteen hips with early stage osteonecrosis diagnosed by bone scintigraphy were studied, as were 22 contralateral normal hips (from unilateral cases) and 22 roentgenographically and scintigraphically normal hips in patients who had been administered corticosteroids. All hips demonstrated abnormal superior retinacular arteries in the extraosseous area, and small arteries penetrated 14 hips with early stage osteonecrosis. Abnormal findings were noted in 17 of 22 contralateral normal hips and in 20 of 22 normal hips with corticosteroid administration. Follow-up roentgenographic analysis showed that the hips with small arterial penetration most often developed osteonecrosis. There were two important findings: (1) The blood supply of the superior retinacular arteries from the extraosseous site was impaired. (2) Revascularization was observed not only in hips with early stage osteonecrosis but also in contralateral normal hips and normal hips with corticosteroid therapy. Osteonecrosis is not necessarily a consequence of a single episode of impairment of blood supply of the femoral head but that of a repetitive episode if interruption of revascularization.


Assessing and diagnosing developmental disorders that are not evident at birth: parental evaluations of intake procedures. Studies of parental satisfaction with the way in which diagnostic information is imparted have generally focused on conditions that are evident from birth. Diagnostic disclosure for disorders in which the onset is later, and less readily identifiable, has received less attention. The present study investigated parents' views of diagnostic procedures for children whose developmental delays only became apparent as they grew older. The degree of satisfaction was related to the type of diagnosis given but 2 weeks after the assessment 87% of parents described themselves as being very or fairly satisfied. However, levels of satisfaction were lower at a later follow-up than they had been initially. Moreover, several specific aspects of the assessment fell short of the ideal model and parents offered a number of practical and cost-effective suggestions as to how these could be remedied. These ranged from ways in which pre-assessment procedures could be made more useful, how diagnostic information should be presented and how the physical setting could be improved.


Relationship of urinary serotonin excretion to cigarette smoking and respiratory symptoms. The Normative Aging Study. The relationship of 2-h urinary excretion of serotonin and 5-hydroxyindoleacetic acid (5-HIAA) to cigarette smoking and respiratory symptoms was examined among 631 male participants in the Normative Aging Study (age range, 44 to 85 years). The amount of serotonin excreted in urine was inversely related to age (p less than 0.001). Mean 2-h excretion of serotonin varied from 8.01 micrograms for men 40 to 49 years of age to 5.84 micrograms for those 70 years of age or over. No clear relationship was evident between the amount of 5-HIAA excreted in urine and age. After adjustment for age, current smokers were found to excrete more serotonin (p less than 0.001) and 5-HIAA (p = 0.001) than never smokers. Former smokers did not differ significantly from never smokers in these respects. After adjustment for age and smoking status in a multivariate model, chronic cough was a significant predictor of serotonin excretion (p = 0.005); chronic cough was less predictive of 5-HIAA excretion (p = 0.07). Other respiratory symptoms were unrelated to urinary excretion of serotonin and 5-HIAA. The mechanisms underlying the observed relationships of urinary serotonin and 5-HIAA excretion to smoking and to chronic cough and their potential relevance to chronic bronchitis remain to be determined.


Interferon gamma-1b. A review of its pharmacology and therapeutic potential in chronic granulomatous disease. Chronic granulomatous disease is a group of rare x-linked or autosomal genetic disorders of the phagocytic NADPH oxidase system involved in host defence against various microorganisms. It is manifested by a common phenotype consisting of recurrent serious, life-threatening infection and granuloma formation. Following the finding that interferon gamma-1b (IFN gamma-1b) can potentiate phagocyte activity in some other disease states as well as restoring defective phagocyte NADPH oxidase system activity in at least some patients with chronic granulomatous disease, a large-scale placebo-controlled trial was undertaken with IFN gamma-1b in patients with chronic granulomatous disease. Long term treatment with a therapeutic dosage of IFN gamma-1b produced a significant reduction in the incidence of serious clinical events necessitating hospitalisation. The relative risk of serious infection and the number of days in hospital were each reduced by about two-thirds, and the mean duration of hospital stay by about one-third in those who did experience infection. The greatest therapeutic benefit was found in patients aged less than 10 years, but all patients were improved regardless of age, sex, use of prophylactic antibiotics or genetic pattern of inheritance. The drug was well tolerated with the commonest adverse effects (e.g. fever, headache, chills, injection site erythema) usually being mild, transient, and relieved by symptomatic treatment. IFN gamma-1b therefore provides an effective and well tolerated therapy for patients with chronic granulomatous disease, offering an important clinical advance in the treatment of this rare genetic disorder by improving the prognosis of its serious and life-threatening infectious sequelae.


Fosinopril. A review of its pharmacodynamic and pharmacokinetic properties, and therapeutic potential in essential hypertension. Fosinopril is a phosphinic acid prodrug which, after oral administration, undergoes rapid hydrolysis to its active diacid, the angiotensin converting enzyme (ACE) inhibitor fosinoprilat. Unlike other ACE inhibitors, fosinoprilat has a compensatory dual route of elimination and is cleared by the liver and kidneys. Thus, in patients with diminished renal function increased hepatic clearance of fosinoprilat is noted and, similarly, in patients with diminished hepatic function increased renal clearance seems to occur. Because of this compensatory elimination, fosinopril therapy for all patients can begin with the same recommended dosage. Fosinopril 10 to 40mg administered once daily is an effective antihypertensive regimen that has shown efficacy similar to that of enalapril 5 to 10 mg/day, propranolol 80 to 160 mg/day, hydrochlorothiazide 25 to 50 mg/day and sustained release nifedipine 40 mg/day in preliminary clinical trials. Generally, fosinopril is well tolerated and adverse events associated with the drug are usually mild and similar to those associated with other ACE inhibitors. Thus, fosinopril appears to be a useful alternative to certain 'established' agents used for treating patients with essential hypertension.


Vasodilators. A re-evaluation of their role in heart failure. Over the past 25 years, the concept of circulation in heart failure has evolved from that of a simple circuit with a weak pump and high pressures to a complex integrated system of cellular modification, cardiac compensation and systemic neurohumoral responses. The original model of cardiac afterload as the systemic vascular resistance has been refined to reflect the interdependence of preload and afterload and the central role of atrioventricular valve regurgitation. It is becoming increasingly apparent that the impact of vasodilator therapy far exceeds the direct haemodynamic effects on preload and afterload, and depends on the mechanism by which vasodilation is achieved, with increasing emphasis on those agents which oppose neurohumoral activation. The potential goals of therapy have broadened to include not only haemodynamic stabilisation through tailored therapy for patients referred with advanced heart failure, but also the prevention of disease progression for patients with asymptomatic ventricular dilation. As the different profiles of heart failure have come to be recognised, the purpose and design of vasodilator treatment must now be considered individually for each patient.


Primary pulmonary hypertension. Practical therapeutic recommendations. Primary pulmonary hypertension (PPH) is a rare disease of unknown aetiology which typically results in right heart failure and death within several years of the onset of symptoms. While there is no cure for PPH, several pharmacological and surgical approaches to treatment have been developed over the past decade which proved useful in a significant proportion of patients. In particular, vasodilator therapy may produce sustained haemodynamic and symptomatic improvement in up to approximately two-thirds of patients; in the remaining patients, vasodilators may either produce no benefit or result in deterioration. The calcium channel blocking agents are the most widely used oral vasodilators; continuous intravenous infusions of epoprostenol (prostacyclin; prostaglandin I2) have been used in some patients who are refractory to oral therapy, particularly as a bridge to transplantation. While combined heart-lung transplantation has been considered the surgical procedure of choice for severe pulmonary hypertension, single lung transplantation has been performed successfully in a small number of patients, and may be the preferred approach in patients with reasonably preserved right heart function.


Pharmacological management of cancer pain. Cancer pain remains a major cause of suffering. Improvements in its management have made unrelieved cancer pain unacceptable. While pharmacotherapy is the mainstay of cancer pain treatment, other options such as radiotherapy, nerve blocks, etc., have to be considered as well. A comprehensive approach must also address psychosocial issues. A successful pharmacotherapy programme for cancer pain requires careful assessment of the origin and cause of the pain. The selection of analgesics has to be rationalised using a sequential approach such as the WHO stepladder. Oral application by the block in an individually titrated dosage is recommended. Although morphine remains the most useful opioid, it should be used in combination with nonopioids. Co-analgesics, which contribute to analgesia without being classical analgesics, should be used to treat pain of specific origin. Here membrane-stabilizers, antidepressants and steroids play an often underestimated role in the treatment of neurogenic pain. Anxiolytics and major tranquillisers should be avoided because they cause sedation without improving quality of analgesia. Calcitonin, diphosphonates and spasmolytics are of minor importance in this regard. Finally, concomitant medication to treat side effects of the therapy may be necessary in formulating a comprehensive treatment plan.


Practical treatment recommendations for the safe use of anaesthetics. General anaesthesia is the reversible depression of central nervous system function. There is still no agreement over what constitutes depth of anaesthesia, and the clinical anaesthetist must thus titrate drug input according to clinical signs (heart rate, blood pressure, somatic movement, autonomic responses). The potency of inhalational agents may be expressed in terms of the MAC (minimum alveolar concentration); comparable end-points (including blood concentrations) have been proposed for the intravenous agents. Kinetic infusion regimens can be constructed for the intravenous agents to achieve the ED95 concentrations required to provide clinically adequate anaesthesia. However, because of individual differences in drug kinetics and dynamics, as well as the influences of disease states and intercurrent therapy, the clinician will titrate the dose according to response. Administration of volatile or intravenous anaesthetics by fixed regimens may result in either overdosage or the risk of patient awareness. The choice of anaesthetic drug is usually based on the nonhypnotic side effects of the different agents--including their central and regional cardiovascular effects, the speed and completeness of recovery, and the need to provide intraoperative analgesia. In addition, special techniques and drugs are often needed for neurosurgical, cardiothoracic and obstetric anaesthesia. All anaesthetic agents (inhalation and intravenous) have other side effects (such as cardiorespiratory depression and organ toxicity related to the liver or kidney). Both halothane and enflurane may be responsible for postoperative hepatic dysfunction, while the metabolism of enflurane can also result in nephrotoxicity in patients with pre-existing renal dysfunction. Isoflurane has been reported to cause 'coronary steal' in patients with ischaemic heart disease through its coronary vasodilator properties.(ABSTRACT TRUNCATED AT 250 WORDS)


Intravenous immune globulins. A review of their uses in selected immunodeficiency and autoimmune diseases. Intravenous immune globulin (IGIV) was introduced a decade ago as a therapy for primary immunodeficiency diseases. It proved to be a valuable therapeutic substance for this purpose and is now considered to be the treatment of choice. The intent was to supply ubiquitous anti-infectious agent antibodies through passive immunisation to replace deficient circulating antibody content. During such therapy, unexpected benefits were noted in thrombocytopenic patients. Since that time, the therapeutic indications for IGIV infusions have greatly increased, with a particular interest in infectious, haematological and autoimmune diseases. This review summarises the status of IGIV therapy in haematological diseases within the categories of primary immunodeficiency diseases, secondary immunodeficiency states and autoimmune syndromes. The majority of firm data have been gathered on the treatment of patients with primary immunodeficiency disease. These data are reviewed from the aspect of anticipated therapeutic response and side effects. Emphasis should be placed on the IgG circulating blood levels as there is a need for individualizing therapy because of marked interindividual patient variation. The use of IGIV therapy in primary and secondary immunodeficiency states should consider the potential benefits to be attained in haematological malignancies and related complications which may be magnified by chemotherapy and radiation therapy. The mode of action of IGIV in autoimmune diseases, although not yet precisely determined, may involve establishing reticuloendothelial blockade or immunomodulation by supplying anti-idiotype antibodies.


Reaction of adrenal glomerular zone to detergents. The effects have been investigated of chronic exposure to detergents on volumetric and histochemical features of the glomerular zone of adrenal glands in the white rat. Neutral lipids, triglycerides, phospholipids and nucleic acids were analysed. There was an increase in the quantities of all the three fatty substances in the corticocytes of the glomerular zone and the quantity of ribonucleic acid was enhanced, whereas that of deoxyribonucleic acid was reduced. The volumes of the nuclei differed significantly between the experimental and the control animals.


Characterization of a cis-acting regulatory element involved in human-aromatase P-450 gene expression. The characteristics of a cis-acting regulatory region involved in the human-aromatase P-450 gene have been examined by transient expression analysis. The region spans from -242 - -166 relative to the cap site of the gene. A fragment containing the region excised from the gene enhances heterologous promoter activity as well as its own promoter activity in a position-independent and orientation-independent manner. The fragment exerts its enhancer activity in human BeWo choriocarcinoma cells in which the aromatase P-450 gene is expressed, but not in other cell lines tested. Deletion of 38 bp from the 3' end of the fragment results in a complete loss of enhancer activity. A gel-retardation assay with nuclear extracts from BeWo cells suggests the existence of a nuclear factor(s) which interacts with the fragment. These results suggest that the regulatory element in the fragment is involved in efficient transcription of the human-aromatase P-450 gene.


The nucleotide and deduced amino acid structures of sheep and pig fetuin. Common structural features of the mammalian fetuin family. This study was initiated to gain further insight into the structural features of the mammalian fetuin family. The cDNA structures of sheep and pig fetuin were determined. The cDNA insert encoding sheep (pig) fetuin comprised 1550 (1470) nucleotides, including 54 (46) nucleotides encoding a signal peptide of 18 (15) residues and 1038 (1041) nucleotides encoding the 346 (347) amino acids of the mature plasma protein. The predicted amino-terminal sequence of the mature pig fetuin was confirmed by the amino-terminal sequence of the purified protein. However, two alternative sheep amino-terminal sequences were found in fetuin purified from the plasma of a single sheep fetus; the minor product was the one predicted by comparison with other fetuin sequences while the major product was two amino acids longer. Comparison of the deduced amino acid sequences of sheep and pig fetuin showed an extensive sequence identity between them (75%) and with other proteins of the mammalian fetuin family, i.e. human alpha 2-HS glycoprotein, and bovine and rat fetuins. Twelve cysteine residues were found at invariant positions in all fetuin sequences, suggesting strongly that the arrangement of disulphide bridges identified in human alpha 2-HS glycoprotein is common to the members of the family. Further sequence comparisons revealed that the structures of mammalian fetuins are organised in three domains: two cystatin-like domains (D1 and D2) and a complex carboxyl-terminal domain (D3). The proposed three-domain structure of the protein is reflected in the organisation of the rat fetuin structural gene which has recently been published.


Diminished capacity to release metabolites of nitric oxide synthase in macrophages loaded with oxidized low-density lipoproteins. Activation of J774-macrophages with lipopolysaccharide (LPS) or LPS and recombinant interferon-gamma (IFN-gamma) induced nitric oxide (NO) synthase activity, as measured by the production of nitrite and citrulline. NO synthase activity was suppressed by loading the cells with oxidatively modified low-density lipoprotein (ox-LDL) but not with acetylated LDL (ac-LDL), although the intracellular lipid accumulation was comparable. This suggests that the extent of activation of lipid-loaded macrophages may be influenced by the type of lipid.


Amiodarone. An overview of its pharmacological properties, and review of its therapeutic use in cardiac arrhythmias. Amiodarone, originally developed over 20 years ago, is a potent antiarrhythmic drug with the actions of all antiarrhythmic drug classes. It has been successfully used in the treatment of symptomatic and life-threatening ventricular arrhythmias and symptomatic supraventricular arrhythmias. In patients with left ventricular dysfunction amiodarone does not usually produce any clinically significant cardiodepression and the drug has relatively high antiarrhythmic efficacy. Preliminary studies indicate that amiodarone may have a beneficial effect on mortality and survival in certain groups of patients with ventricular arrhythmias, an action probably related to both its antiarrhythmic and antifibrillatory effects. The adverse effect profile of amiodarone is diverse, involving the cardiac, thyroid, pulmonary, hepatic, gastrointestinal, ocular, neurological and dermatological systems. Interstitial pneumonitis and hepatitis are potentially fatal, but the vast majority of adverse events are less serious, and some may be dose dependent. Pretreatment monitoring, regular assessments and the use of minimum effective doses are, therefore, necessary. Thus, with appropriate monitoring to control its well recognised adverse effects amiodarone has an important place as an effective 'broad spectrum' antiarrhythmic drug which has, so far, been used when other treatments have proved ineffective. More recent preliminary data also suggest that it may also have a beneficial effect in the prevention of sudden death in some patients.


Endopeptidase 24.15 modulates bradykinin-induced contraction in guinea-pig trachea. Guinea-pig tracheal strips were contracted with cumulative concentrations of bradykinin or substance P in the presence of thiorphan, an inhibitor of endopeptidase 24.11 and of angiotensin-converting enzyme, or in the presence of N-[1-(R,S)-carboxy-3-phenylpropyl]Ala-Ala-Phe-para-aminobenzoate (cFP-AAF-pAB), a selective inhibitor of endopeptidase 24.15. The concentration-effect curve of bradykinin was shifted to the left in the presence of each inhibitor whereas the curve of substance P was sensitive to thiorphan but not to cFP-AAF-pAB. These results show that endopeptidase 24.15 may modulate the contractile effect of bradykinin but not that of substance P in the guinea-pig trachea.


Differential sensitivity of CG and CCG DNA sequences to ethionine-induced hypomethylation of the Nicotiana tabacum genome. Plant DNA is distinguished from the DNA of all other organisms by its high content of 5-methylcytosine (5mC). 5mC levels may amount to 30% of total cytosines, distributed between the sequences CG and CXG. The results presented here show that the methylation status of CXG sequences could be influenced by culturing tobacco tissues on subtoxic concentrations of ethionine. The hypomethylating effect of ethionine, evaluated as the capability of MspI or HpaII to cleave the DNA, proved to be rather specific for CCG and differed from that of 5-azacytidine which did not discriminate between CG and CXG sequences.