Atopic disease in ulcerative colitis and Crohn's disease.

Three hundred patients with ulcerative colitis, 200 with Crohn's disease and matched control subjects completed questionnaires about atopic disease. They were asked whether they had ever suffered from asthma, hay fever, allergic rhinitis or eczema; in ulcerative colitis all of these features occurred with twice the frequency, but in Crohn's disease only eczema was more common than in controls.


'Barn allergy': asthma and rhinitis due to storage mites.

The differential diagnosis of asthma in farm workers is between a type I hypersensitivity to a pollen or dust allergen and a type III hypersensitivity to mould antigens. We describe asthma and rhinitis, and not farmer's lung, in a farming population due to the non-pyroglyphid grain storage mite. Before a diagnosis of farmers' lung due to mouldy hay is made in any patient whether or not precipitins to Micropolyspora faeni are present, skin tests for storage mite should be made. If these are positive a diagnosis of 'barn allergy' should be considered and a trial of sodium cromoglycate be given.


What kind of thing is 'being clumsy'?

The term 'clumsy' is used both as a colloquial pejorative and as a specific diagnostic entity. The meaning of the word and the possible means by which a child may come to be regarded as clumsy are discussed. The implications for management are considered.


Mite allergen content in commercial extracts and in bed dust determined by radioallergosorbent tests.

Radioallergosorbent (RAST) direct binding and inhibition type assays were used to quantitate the mite (Dermatophagoides pteronyssinus) allergen content of four commercial mite extracts and a laboratory prepared extract from freeze-dried mites. The content of mite allergen in extracts prepared from twenty samples of dust vacuumed from bedding was measured by RAST inhibition assay. The four commercial mite extracts designated A, B, C and D, and the laboratory extract, designated L, contained 52, 265, 108, 1.5 and 581 arbitrary units of allergen/ml for the direct binding assay and 128, 111, 217, approximately 1 and 1083 arbitrary, but different, units of allergen/ml for the inhibition assay respectively. Qualitative differences between at least two extracts were suggested by the different slopes obtained when allergen binding of anti IgE was plotted against the volume of extract used in the direct binding assay. Differences in slope between the two extracts were also apparent when they were used in the inhibition assay. The quantities of mite allergen/gm of bed dust expressed in arbitrary units for the inhibition assay were 24 to 457 (mean 129) units. These quantities are similar to and sometimes greater than the quantity in 1 ml of mite extract and so confirm bed dust as a potent source of mite allergen. There was no significant correlation between the weight of dust, the numbers of dead and live mites and the allergen content of dust.


Action research and evaluation: can research and practice be successfully combined?

Educative intervention with the young severely subnormal child and his family is envisaged as an example of 'action research' in which the dual roles of the practitioner-researcher and the client-subject are examined. A view of the effect of the interaction of these roles on the intervention and its evaluation is given, and conclusions drawn concerning the nature of evaluation appropriate in future projects of this kind.


Living in one-parent families: attainment at school.

Data drawn from the National Child Development Study enabled an examination to be made of the school attainment of children who had spent any of their childhood in a one-parent family. Overall, children in one-parent families had lower scores on tests of attainment, but after adjusting for background factors this was no longer the case for the reading test, and the differences on the mathematics test were reduced to non-significance. An examination of particular aspects of these background factors showed that the low income of many of the one-parent families was a substantial part of their social disadvantage associated with low attainment, but this was not the case for their housing conditions. Comparisons within the group of one-parent families revealed little variation. This meant that there was no evidence that either the age at which the child's family broke up nor the reason for the break up (i.e. divorce or widowhood) was important in terms of school performance, although there was a very slight indication that children who had acquired a substitute parent figure had lower test results than those whose parents had remained alone.


Intravenous NAB 365 (clenbuterol) and terbutaline in exercise-induced bronchospasm (EIB).

In a single-blind, placebo-controlled, cross-over study, the effects of intravenous 20 micrograms NAB 365 (clenbuterol) and intravenous 250 microgram terbutaline were compared in nine asthmatic patients with bronchospasm induced by exercise. The exercise consisted of treadmill running. The two beta-adrenergic bronchodilator drugs effectively prevented post-exercise bronchospasm in eight of the nine patients. The effects were superior to those in the untreated group and in the placebo-treated group. Six of the patients treated with terbutaline complained of adverse reactions (tremor, palpitation, tachycardia). No adverse reactions were observed after clenbuterol.


Analysis of the non-immunological activity of allergen extracts in cutaneous tests.

The technique proposed by the National Institute for Biological Standards and Control, London (NIBSC, 1977) for the control of allergenic extracts for non-immunological activity in skin tests has proved to be both sensitive and useful. We have observed 'false positive' results to substances such as merthiolate or glycerine and also to lyophilised preparations with sugar, notably glucose, as the carrier substance. Other factors studied in 'false positive' reactions to a given extract were pH, osmotic pressure and the presence of traces of endotoxins or histamine. Individually none of these provided satisfactory explanations for the false positive reactions and the use of antagonists or inhibitors of the release of chemical mediators only confirmed the complexity of the non-immunological reaction and the participation of factors other than histamine.


Asthma, aspergilloma and anergy in a patient being treated for pulmonary tuberculosis.

A twenty-one-year old white girl developed cavitating pulmonary tuberculosis which was complicated by nocturnal asthma, aspergilloma and anergy. In addition she developed systemic symptoms suggestive of a hypersensitivity reaction to the aspergilloma.


Increased concentrations of free fatty acids and glycerol and altered creatine kinase MM isoenzyme patterns in certain diabetic patients.

We report unusually high concentrations of free fatty acids and glycerol in sera of patients with adult-onset diabetes, and the accompanying alterations in creatine kinase isoenzyme MM patterns associated with such patients. The serum samples with increased free fatty acids also showed increased electrophoretic movement of the MM isoenzyme on cellulose acetate membranes. Fatty acid concentrations found in such samples averaged 9.88 +/- 5.65 (SD) meq/L and the average glycerol concentration was 153 +/- 115 (SD) mg/L. The serum glycerol concentrations correlated with those of the free fatty acids (r = 0.886, slope = 0.309, intercept = 4.76).


Concentrations of ionic, total, and bound fluoride in plasma.

We found no significant difference between the means for ionic, bound, and total fluoride concentrations in the plasma of male and female subjects of the same age, living in a community with fluoridated water. When results for the 264 fasting subjects were therefore combined according to age, they indicated that persons over 60 years of age have a significantly higher mean ionic (3.89 mumol/L) and total (6.58 mumol/L) fluoride concentration in plasma than do younger age groups. For younger age groups, means ranged from 2.74 to 3.05 mumol/L for ionic fluoride and from 4.74 to 5.58 mumol/L for total. The bound fluoride concentration was lower in individuals 21 to 30 years of age (1.89 mumol/L) than in older age groups (for whom means ranged from 2.42 to 2.68 mumol/L), but was not significantly different from that of individuals who were younger (2.21 mumol/L). A tendency for the mean ionic fluoride concentration to increase with age was noted, but the concentration was significantly higher than the preceding decade group only in those persons over 60 years of age.


Immunochemical determination of human immunoglobulins: use of kinetic turbidimetry and a 36-place centrifugal analyzer.

We describe the measurement of human immunoglobulins IgG, IgA, and IgM in diluted sera, with diluted commercial monospecific antisera, with use of a 36-place centrifugal analyzer (RotoChem IIa; American Instrument Co., Silver Spring, MD 20920). The assay involves a two-point kinetic turbidimetric technique. The changes in absorbance at 340 nm between 10 and 255 s are taken for standard curve construction with use of a computer-generated cubic least-squares fit. Patients' samples are quickly calculated from the stored curve. Multiple time-interval readings are taken to observe the reaction kinetics. The lowest detectable concentrations are: IgG, 5 mg/L; IgA, 20 mg/L; and IgM, 20 mg/L. Correlation with radial immunodiffusion was excellent. Precision, accuracy, linearity, and sensitivity were very acceptable. Antigen or antibody excess can be easily detected. The accuracy of the proposed method when measuring idiotypic monoclonal proteins is greater than radial immunodiffusion. Up to 30 patients' samples can be analyzed at one time, and calculation of test results by use of the computer program is efficient and rapid.


Preservation, distribution, and assay of glucose in blood, with special reference to the newborn.

Of several preservatives we tested, a mixture of "isotonic" sodium fluoride and sodium iodoacetate (17.7 and 5.0 g/L, respectively) best inhibits glucose loss from blood collected from the newborn. However, the inhibition is incomplete. Twenty microliters of blood is diluted with 100 microliter of preserving solution. After centrifugation, the glucose in 100 microliter of prewarmed supernate (diluted plasma) is measured in a Beckman Glucose Analyzer, based on the action of a glucose reagent and the rate of oxygen consumption. The method, in effect, determines the glucose in whole blood, except for about 5% of the total that remains in the erythrocytes. Glucose is distributed in blood according to the water content of cells and plasma. From concentrations found in cells and plasma, we can calculate the value for glucose in whole blood with an accuracy of 95.2%. The modified method we describe shows acceptable precision, as judged from within-run and day-to-day CV--generally well below 5% for each.


Further observations on the incidence and nature of atypical creatine kinase activity.

The interference of atypical creatine kinase (CK; EC 2.7.3.2) with anion-exchange methods for the measurement of the CK-MB isoenzyme is now firmly established. False-positive results from this source are much more common than interferences caused by the BB isoenzyme. Atypical CK, at least in some patients, does not appear to be a genetic variant, nor could we relate it to a specific clinical diagnosis, to hypoxia, or to administration of a particular drug. Its behavior in an immuno-inhibition test for CK-B subunit indicates an immunological difference from the normal M subunit. Thus the change in immunological properties is associated with the altered electrophoretic properties. The variability of electrophoretic patterns suggests that atypical CK may represent multiple forms of creatine kinase rather than a unique entity.


Radioimmunoassay of 17 alpha-hydroxyprogesterone in saliva, parotid fluid, and plasma of congenital adrenal hyperplasia patients.

We report a radioimmunoassay sensitive enough to determine 17 alpha-hydroxyprogesterone concentrations in 200 microliter of parotid fluid or mixed whole saliva. Because the correlation of concentrations in matched samples of parotid fluid and saliva was excellent (r = 0.98), we exclusively used saliva, which is easier to collect, in later studies. The assay is specific; saliva samples assayed with and without thin-layer chromatographic purification showed no significant difference. The assay is also precise, and has a lower limit of sensitivity of 4 pg per assay tube. In 14 patients having congenital adrenal hyperplasia from a C21-hydroxylase enzyme deficiency, all of whom were receiving cortisol replacement therapy, the range in 17 alpha-hydroxyprogesterone concentrations observed in saliva (67-26,300 pmol/L) was about 20-fold that seen in 32 healthy children (90-1520 pmol/L). The close correlation (r = 0.91) between 17 alpha-hydroxyprogesterone concentrations in matched samples of saliva and plasma from these patients indicates that determination of steroids in saliva could well replace determination in plasma. This concept is supported by 17 alpha-hydroxyprogesterone concentrations monitored throughout 24 h from one patient and following stimulation with synthetic corticotropin in another patient.


A solid-phase enzyme-linked immunosorbent assay for the quantitation of human plasma alpha 1-acid glycoprotein.

We describe a solid-phase enzyme-linked immunosorbent assay for alpha 1-acid glycoprotein in human plasma. Plasma samples are incubated with alkaline phosphatase-linked, purified alpha 1-acid glycoprotein in alpha 1-acid glycoprotein-specific antibody-coated polystyrene tubes. The alkaline phosphatase that becomes attached to the tube via an immunological reaction between the alpha 1-acid glycoprotein and the specific antibody is measured spectrophotometrically. This assay is accurate reproducible, simple, and economical. As little as 4 microgram of alpha 1-acid glycoprotein per liter can be detected. The normal range for alpha 1-acid glycoprotein in the plasma of healthy adults, as measured by this method, is 0.48-1.27 g/L; the range is significantly different, 0.29-0.73 g/L, for women who are taking oral contraceptive pills.


Measurement of aspartate aminotransferase activity: effects of oxamate.

Oxamate, a potent inhibitor of lactate dehydrogenase, is shown also to inhibit aspartate aminotransferase activity, both in human serum and in purified isoenzymes of human origin. The inhibition was competitive with respect to 2-oxoglutarate for both isoenzymes. The apparent Ki was 29 mmol/L for the cytoplasmic enzyme and 17 mmol/L for the mitochondrial enzyme. Noncompetitive inhibition was found between oxamate and aspartate. At saturating concentrations of substrate (2-oxoglutarate greater than or equal to 15 mmol/L, L-aspartate greater than or equal 150 mmol/L) oxamate inhibited the mitochondrial enzyme but had less effect on the cytoplasmic isoenzyme. Oxamate at 40 mmol/L inhibited the enzyme in serum by 11 and 9% in assays containing 2-oxoglutarate at 6.7 and 15 mmol/L, respectively. This concentration of oxamate inhibited enzyme activity in serum by 5% more than did the same concentration of Cl- (itself an inhibitor). Oxamate (less than or equal to 30 mmol/L) had no measurable effect on the stability or activity of porcine malate dehydrogenase. Until the effects of its inhibitory properties are considered, addition of oxamate to suppress lactate dehydrogenase-mediated side reactions in the assay of aspartate aminotransferase cannot be recommended.


Simplified determination of urinary delta-aminolevulinic acid in a wide range of concentrations.

We describe a simple method for measuring delta-aminole-vulinic acid in urine without the need for the seldom-used but time-consuming internal-standard addition step. It combines the analytical procedure described by Tomokuni and Ogata [Clin. Chem. 18, 1534 (1972)] and a correction based on urine density to obtain, more rapidly and less expensively, results as accurate as those given by the Davis and Andelman [Arch. Environ. Health 15, 53 (1967)] chromatographic method.


Assay of 1,25-dihydroxy vitamin D3 by isotope dilution--mass fragmentography.

We describe a specific assay of 1,25-dihydroxy vitamin D3 in human serum, based on isotope dilution-mass fragmentography. We added [26-2H3]-1,25-dihydroxy vitamin D3 to a fixed amount of serum. The steroids were extracted with chloroform/methanol and purified by liquid chromatography. The purified materal was converted into the trimethylsilyl ether and analyzed. Unlabeled 1,25-dihydroxy vitamin D3 was quantitated from the ratio between the tracings at m/e 452 and 455. The two ions used correspond to loss of two trimethylsilyloxo functions from the molecular ion. Essentially the same results, with some problems of interference, were obtained when we used the more intense ions at m/e 131 and 134 (corresponding to cleavage between C-24 and C-25). The detection limit was about 5 ng/L of serum; the coefficient of variation was about 6%. The accuracy of the method was assessed by recovery experiments. 1,25-Dihydroxy vitamin D3 in sera from 15 healthy subjects was found to average 55 +/- 10 ng/L(+/- SD). We believe this represents the first determination of 1,25-dihydroxy vitamin D3 in serum by use of a method not based on radioimmunoassay or receptor assay. Slightly lower values have been reported by those latter techniques.


New fluorometric analysis for mandelic and phenylglyoxylic acids in urine as an index to styrene exposure.

I describe a new fluorometric method for determination of mandelic and phenylglyoxylic acids in urine, the fluorometry being preceded by extraction into ether and thin-layer chromatography. The chromatographically separated acids were quantitated after conversion to stable highly fluorescent derivatives by treatment with concentrated sulfuric acid. This method proves to be more precise, accurate, and reproducible than the existing colorimetric method. The limit of detection is 2 microgram of either acid per milliliter of urine with a CV of less than 15%. The standard curve for either acid is essentially linear from 2 to 100 microgram/mL of urine, with a correlation coefficient (r) of 0.97. No satisfactory correlation was obtained between the concentrations of either acid metabolite as measured in rat urine by fluorometry and colorimetry. The present method is considered suitable for routine biological monitoring of persons exposed to both high and low concentrations of styrene.


Separation of protoporphyrins and related compounds by reversed-phase liquid chromatography.

We describe the separation of protoporphyrin and related porphyrins by reversed-phase "high-performance" liquid chromatography, with fluorometric detection. We used the method to demonstrate that acid hydrolysis of the dimethyl ester of protoporphyrin IX is complete in 2 to 3 h and is followed by the acid-catalyzed conversion of protoporphyrin IX to a chemical species that chromatographic evidence indicates to be hematoporphyrin IX. In addition, the method was used to evaluate the purity of a commercial preparation of protoporphyrin IX and was also demonstrated to have the sensitivity and specificity needed for measuring the protoporphyrin IX content of whole-blood.


Improved automated kinetic determination of uric acid in serum by use of uricase/catalase/aldehyde dehydrogenase.

The enzymatic determination of serum uric acid by use of uricase, catalase, and aldehyde dehydrogenase according to Haeckel [J. Clin. Chem. Clin Biochem. 14, 101 (1976)] showed interferences from ethanol-converting enzymes, which are present in some patients' sera. We have identified these enzymes as alcohol dehydrogenase isoenzymes. Among other substances, a mixture of pyrazole and oxalate can be used to eliminate these interferences. This inhibitor system gives good results when used in the automated kinetic uric acid determination, as is shown by a comparison with the manual assay for uric acid according to Kageyama [Clin. Chim. Acta 31, 421 (1971)].


Preparation of cytosol for steroid receptor determinations with the Beckman "Airfuge".

Preparation of cytosols from several estrogen target organs was compared, with use of the Beckman Airfuge and a preparative ultracentrifuge. The specific estrogen-binding capacities of cytosols prepared with these instruments were indistinguishable. Similarly, the sedimentation profiles of estrogen-receptor species as determined by sucrose gradient centrifugation were identical. We conclude that the Airfuge provides a rapid, accurate, and inexpensive means of preparing cytosols for measurements of steroid receptors in the clinical laboratory.


Effect of light and temperature on the stability of creatine kinase in human sera and controls.

Commercial control preparations were exposed to light at room temperature for as long as 24 h. Decreases in creatine kinase activity in seven of the nine controls ranged from 28.8 to 83.3%. When these controls were protected from light and stored at various temperatures, the decrease in activity was either eliminated or substantially reduced, indicating that the decreased activity in the presence of light was not due entirely to thermal inactivation. In the absence of oxygen, light had little effect on creatine kinase suggesting that light inactivation is a light-catalyzed oxidative process. The decreased activity observed when some of the controls were exposed to light was not completely restored by incubating with dithiothreitol before analysis. However, increases in creatine kinase activity ranging from about 37 to 176% were observed when freshly prepared controls were pre-incubated with dithiothreitol. Neither light nor dithiothreitol had any effect on the creatine kinase activity in two of the nine controls. When human sera were exposed to light for 24 h, the largest decrease in activity was about 15%. Incubation of fresh human sera with dithiothreitol before analysis caused an average increase in activity of approximately 10%.


Potential clinical applications of derivative and wavelength-modulation spectrometry.

Absorption spectrophotometers are now becoming available that can record the first and second derivatives of absorbance with respect to wavelength. The advantages of this technique and its potential application to clinical analysis are discussed.


Microencapsulated antibodies in radioimmunoassay. II. Determination of free thyroxine.

We describe a method for directly measuring free thyroxine in human serum. Antibody to thyroxine was encapsulated within a semipermeable nylon membrane that excludes substances of relative molecular mass greater than 20,000. The microencapsulated antibody was then presaturated with [125I]thyroxine. Serum incubated with the microcapsules initiated a displacement reaction between free thyroxine and [125I]thyroxine bound to the antibody. The displaced thyroxine was separated from the bound thyroxine by centrifugation and the concentration of free thyroxine determined from a standard curve prepared by use of known amounts of free thyroxine. The within-day coefficient of variation for two control samples was 8.8 and 6.8%; day-to-day precision for the same material was 12.5 and 13.5%. Lipemia, icterus, or hemoglobin had no adverse effect. Interlaboratory evaluation of the procedure revealed no significant difference in mean values for 20 speciments (p greater than 0.05).


Plasma polyamines determined by negative-ion chemical ionization/mass spectrometry.

We have developed an accurate and highly sensitive gas-chromatographic/mass-spectrometric procedure for determining di- and polyamines (putrescine, spermidine, and spermine) in plasma and erythrocytes. Deuterium-labeled analogs of putrescine, spermidine, and spermine were synthesized for use as internal standards. Trifluoroacetyl derivatives of the polyamines were formed during the procedure and then detected with negative-ion chemical ionization/mass spectrometry in combination with multiple ion monitoring. Limits of sensitivity ranged from 0.25 to 1.0 pmol of analyte injected into the instrument.


A kinetic method for glucose that is insensitive to variations in temperature and enzyme activity.

We have adapted for glucose determination a new approach to kinetic analyses [Anal. Chem. 50, 1611 (1978)]; it is 50-fold less dependent upon some experimental variables than is a more conventional rate method. Modification of a commercially available hexokinase/glucose-6-phosphate dehydrogenase reagent system for glucose provides that the rate of production of NADH be first-order in total glucose concentration within about 30 s after sample and reagent are mixed. In the kinetic method, absorbance vs. time data recorded after 30 s and a multiple-linear-regression program are used to compute the absorbance change that would occur if the reaction were monitored to completion. Results demonstrate a linear relationship between glucose concentration and computed absorbance change. Application of the method to 51 human sera without rigorous control of either temperature or reagent composition yielded a regression equation of y = 1.01x -0.3 when kinetic results (y) were compared with equilibrium results (x) for the same samples analyzed in a hospital laboratory.


New developments in analysis of isoenzymes separated by "high-performance" liquid chromatography.

We have developed two enzyme analyzers for use in "high-performance" liquid chromatography. In both systems two detectors are used, placed after the column effluent has been combined with assay reagent. In one system, an absorbance detector is placed before and after a post-column reaction coil. Peaks observed at one detector are subtracted from those at the other, to produce a two-point measurement of enzyme activity. The linear dynamic range was 17--1700 U/L for lactate dehydrogenase (EC 1.1.1.27). In the other system, two reaction coils were used and a single fluorescence detector was placed at the end of each coil. These coils were kept at different temperatures, and an automated switching valve diverted equal amounts of column effluent and reagent into both coils. The fluorescence readings were then subtracted to produce a differential measurement of enzyme activity. The linear dynamic range was 20--1000 U/L. We used both systems to chromatographically analyze lactate dehydrogenase isoenzymes, and could separately determine both the distribution and activity of sample isoenzymes.


Rapid analysis of bilirubin in neonatal serum. I. The binding of bilirubin to albumin.

Evaulation of the severity of jaundice in the neonate may be determined by measuring the reserve binding capacity of serum proteins for free bilirubin. Determination of protein-bound bilirubin has been labor intensive, necessitating multiple runs on gel-permeation chromatography columns or, more recently, enzyme assays or fluorescence quenching analysis. We present a method for quantitation of free bilirubin and of bilirubin-binding capacity of serum by liquid chromatography. A gel-permeation column binds free bilirubin while allowing passage and quantitation of protein-bound bilirubin. Subsequent injection of a desorbing agent releases the adsorbed bilirubin from the column, permitting quantitation of free bilirubin. Bound and free serum bilirubin may be determined directly in less than 15 min using 10 microL of serum. The binding of bilirubin to neonatal serum is seen to be quite different from the binding to adult serum. Ion-exchange chromatography of adult and neonatal serum samples shows that their protein profiles are radically different. This difference probably accounts for the binding characteristics.


Improved method for detecting hemagglutination by centrifugal analysis.

Centrifugal analysis can be used to detect hepatitis B surface antigen, antibody to rubella virus, and fibrin-related antigen. The procedure is performed with the same reagents used in conventional hemagglutination studies. Positive and negative reactions are distinguished by the rates of erythrocyte clearance in the centrifugal field (delta A/delta time); positive cells move more rapidly than negative cells, and this difference varies directly with the concentration of detectable antigen or antibody. This phenomenon is thought to be a result of the greater adhesion of negative cells to the cuvette's surface. Sensitivity and specificity are greater in the centrifugal analysis technique than in the more conventional hemagglutination tests. False-positive reactions are eliminated and the quantitative data are accurate and reproducible.


High-performance liquid chromatography for ultraviolet-absorbing constituents in human urine.

Several liquid-chromatographic systems involving anion-exchange resins and linear acetate gradient elution have been reported previously for the analysis of ultraviolet-absorbing constituents in human urine. We describe a high-performance liquid-chromatographic system on which a macroreticular anion-exchange resin, and stepwise elution are used. Separations were more rapid with ammonium chloride/acetonitrile stepwise elution than with acetate stepwise elution, and urine constituents could be detected at 225 nm with the former system. Analytical conditions were convenient and the method has proven to be useful for routine clinical analysis.


Instrumentation and techniques for analysis of hydrogen peroxide and peroxide-producing reactions involving earthworm (Diplocardia longa) bioluminescence.

Earthworm bioluminescence (species Diplocardia longa) is a sensitive tool for determination of peroxide titers in biological systems, with linearity of the standard curve for hydrogen peroxide extending from 10 nmol/L in the assay to greater than 1 mmol/L, and with sensitivity to 2 pmol of peroxide per assay. This assay's insensitivity to extraneous protein, turbidity, and solutes that are not redox active makes it particularly useful for automated or crude sample analysis and for analysis of oxidases and their substrates in coupled reactions. Examples of this latter application we discuss are analyses of glucose and putrescine oxidases and their substrates. In each case linear standard curves are obtained, covering several decades of concentration. The coupled reactions also allow in situ detection of oxidases in electrophoresis gels. We also describe a small, portable photometer system and an on-line computer-controlled instrument system that has facilitated development of these assays and will also allow automatic scanning of bioluminescence in gels. The data-collection and decision-making algorithms of this system are simple and easily adaptable for use in an inexpensive computer-based photometer system.


Flow-injection analysis with chemiluminescence detection.

In "flow-injection analysis," the sample is inserted into a stream of reagent by use of a sample-injection valve. Mixing occurs downstream from the valve in a coil of tubing. With chemiluminescence detection this coil is positioned in front of a photomultiplier. We have evaluated the system for detection of hydrogen peroxide, using the luminol reaction with cupric ion as a catalyst. The effects of flow rate, sample volume, and reaction kinetics on the magnitude, duration, and repeatability of the chemiluminescent response have been evaluated. Precisions of 1 to 2% relative standard deviation on replicate measurements are readily achievable. A sample throughput of six samples per minute is possible with very little peak overlap. This detection system can be coupled to any process in which peroxide is generated.


Bioluminescence assay of creatine kinase and its isoenzymes in serum and cerebrospinal fluid.

We examined the sensitivity of bioluminescence for the determination of very low concentrations of creatine kinase brain-type subunit (CK-BB) in serum and in cerebrospinal fluid. To optimize the sensitivity of CK-isoenzyme assays and eliminate possible sources of error, we separated the isoenzyme fractions by using inhibiting anti-MM and precipitating anti-MM and anti-BB antibodies. The results with the bioluminescence assay correlated with spectrophotometric values such that r = 0.97 for the total CK activity and r = 0.98 for the CK-B activity. The reproducibility of the present method was comparable with the spectrophotometric method and was even better at low enzyme activities. The within-series precision for assay of total CK activity at 2 U/L corresponded to a CV of 9%; at 13 U/L the CV was 5.8%. All the assays were carried out at 25 degrees C. Even at this low temperature, CK activities as low as 0.2 U/L could be determined. In eight patients without any evidence of cerebral cell damage, total CK activity in cerebrospinal fluid was x = 1.05 +/- 0.6 U/L, and CK-BB activity was x = 0.7 +/- 0.4 U/L. In sera of these patients CK-BB activity was x = 0.6 +/- 0.5 U/L. Differences in CK and CK-BB activities in four patients with transient or progressive brain-cell damage are discussed.


A new screening system for nonsteroidal anti-inflammatory drugs based upon inhibition of chemiluminescence produced from human cells (granulocytes).

A new screening system for nonsteroidal anti-inflammatory drugs that involves use of human phagocytic cells has been developed in which chemiluminescence measurement is used. Luminol-dependent chemiluminescence is measured after the addition of opsonized (coated with antibodies and complement) zymosan particles to human granulocytic leukocytes in the presence or absence of drugs. Of all the compounds tested, indomethacin was the most potent in blocking chemiluminescence, with measurable inhibitory activity at 5 mumol/L. The order of inhibitory potency at 0.1 mmol/L and in the presence of Ca2+ and Mg2+ was indomethacin greater than sodium salicylate greater than fenoprofen Ca greater than tolmetin greater than naproxen greater than ibuprofen. It is likely that the active compound itself must be added to the system because aspirin did not inhibit chemiluminescence, whereas its metabolite, sodium salicylate, was markedly inhibitory. Dexamethasone and methylprednisolone also did not inhibit chemiluminescence. The drugs that inhibit chemiluminescence are also known inhibitors of prostaglandin synthase (cyclooxygenase portion).


Sulfonamide inhibition of human alkaline phosphatase.

Several sulfonamides used as antibacterial or diuretic drugs are potent alkaline phosphatase inhibitors. The mechanism of inhibition may involve binding of the zinc in the active site of the enzyme by the unbonded electron pair on the sulfonamide group nitrogen atom as well as binding of the drug to a second site. Addition of progressively larger groups to this nitrogen leads to an increasing loss of inhibitory capability. Isoenzymes from human liver, bone, kidney, granulation tissue and intestine are inhibited to a similar extent while the placenta isoenzyme is more resistant. It is suggested that some pharmacologic actions of sulfonamides may be due to inhibition of alkaline phosphatase, rather than carbonic anhydrase.


Gelatin derivatives in blood: interference with the determination of inorganic phosphate in serum.

The method of Goldenberg and Fernandez (1966) Clin. Chem. 12, 871--882) for the determination of inorganic phosphate in serum is significantly influenced by turbidity after intravenous application of gelatin derivatives, even at low quantities. This turbidity can be eliminated by the addition of sodium dodecyl sulphate to the reaction solution.


The occurrence of gamma-glutamylphenylalanine in the urine of newborn phenylketonurics.

The urine of untreated phenylketonurics in the first weeks of life contains gamma-glutamylphenylalanine (0.07--0.69 mmol/g creatinine, 12 samples) visible on the normal 2-dimensional electrophoreto-chromatogram. This compound is less prominent or absent when urine from older untreated phenylketonurics is examined and is not seen in normal urine.


The rapid extraction of paraquat from plasma using an ion-pairing technique.

A method for the quantitative extraction of paraquat from plasma is described. The method relies on the use of an organic-soluble ion pair of paraquat with dodecyl sulphate. The use of the extraction procedure in conjunction with the dithionite colour reaction gives a method which is suitable for emergency estimations with a sensitivity limit of 50 micrograms/l. The extraction procedure alone offers a new first step in the clean up of paraquat for analysis by other methods.


Emergency estimations of paraquat in plasma. A comparison of the RIA and ion pair/colorimetric methods.

A comparison was made between the RIA method for paraquat in plasma, and a modified colorimetric method involving ion-pair extraction of paraquat. The comparison was carried out on plasma from patients admitted with paraquat poisoning. The correlation between the methods was good and both were found to be sufficiently rapid for emergency use.


Osmotic stability of erythrocytes in human muscular dystrophy before and after phospholipase treatment.

The stability of washed erythrocytes from patients with muscular dystrophy was determined in hypotonic phosphate buffered sodium chloride. Control cells were more stable than cells from Duchenne and myotonic patients. After pretreatment of the cells with phospholipase from pancreas, snake venom or bee venom in the presence of 14 mmol/l Ca2+, the order of osmotic stability in the 3 groups was not changed. In isotonic phosphate buffered NaCl, however, the erythrocytes of the myotonic patients were much more stable than the cells of the Duchenne and the control group. The lytic process was further studied in control cells with pancreatic phospholipase. 21 +/- 3 (S.E.M.) % of the cells were lysed. This process was (partly) prevented by omitting the phospholipase, by replacement of Na+ by K+ or Li+, by lowering the Ca2+ concentration, by omitting phosphate, by ouabain, by glucose, by ribose, by sucrose, by tetrodotoxin, a Na+-transport inhibitor. Blocking of the Ca2+ transport by La3+ or mersalyl, greatly stimulated the lytic process.


Serum tetraiodothyroacetate (T4A) levels in normal healthy euthyroid individuals determined by gas chromatography-mass fragmentography (GC-MF).

A gas chromatography-mass fragmentography (GC-MF) assay for T4A in human serum is described. The assay involves a multistage extraction, followed by conversion of T4A to its dimethyl derivative prior to GC-MF. Using this assay, the concentration of T4A in 37 normal euthyroid subjects was, mean +/- S.D., 114.5 +/- 26.4 pmol/l, which is considerably lower than previous reports. Thyroxine binding prealbumin (TBPA) did not appear to be a major determinant of T4A concentration.


Determination of volatile fatty acids in urine by gas chromatography on a new stationary phase.

A rapid gas chromatographic method for determination of volatile fatty acids in urine is described. Acids up to five carbon atoms, including their isomers can be separated in less than six minutes on the new stationary phase Fluorad FC430. Good separation and sensitivity is obtained with isothermal conditions.


Catalase, superoxide dismutase, glutathione reductase and thiobarbituric acid-reactive products in normal and dystrophic human muscle.

The level of thiobarbituric acid-reactive products and the specific activities of catalase and glutathione reductase were significantly higher in muscles from patients with major forms of muscular dystrophies over those of control subjects. Superoxide dismutase activity was not altered in dystrophic muscles. These findings indicate the occurrence in human dystrophic muscles of an increased level of lipid peroxidation and the possible activation of certain enzymes that could conceivably inhibit lipid peroxidation in vivo.


Relationship of thyroid status and serum N-acetyl-beta-glucosaminidase isoenzyme activities in humans.

Serum from 28 hyperthyroid, hypothyroid, and euthyroid pre- or early puberty females was examined for N-acetyl-beta-glucosaminidase (HEX) activity. Total, isoenzyme A (labile), and isoenzyme B (stable) levels were determined for this enzyme. A high degree of correlation (r = 0.76; p less than 0.001) exists between total hexosaminidase activity and thyroid hormone levels (as reflected by the Free Thyroxine Index). Examining each isoenzyme individually, A is selectively enhanced (r = 0.84; p less than 0.0005) whereas B displays no significant change regardless of thyroid activity. In hyperthyroid individuals, levels of total hexosaminidase (730 +/- 67 units) (mean +/- S.D.) and isoenzyme A (516 +/- 46) were significantly higher than levels found in either the hypothyroid (total: 547 +/- 30; isoenzyme A: 352 +/- 31) or euthyroid (total: 620 +/- 81; isoenzyme A: 423 +/- 45) groups. However, no change was observed in levels of isoenzyme B among hypothyroid (195 +/- 19), euthyroid (197 +/- 43) and hyperthyroid (215 +/- 32) groups. These data substantiate our earlier findings in the rat, wherein thyroxine administration evoked a similar response in the liver. They are of particular interest in light of the deficiency of HEX A in variants of GM2 gangliosidosis.


Use of small samples of urine to monitor gonadotrophins in menopausal women.

The increased gonadotrophin secretion which occurs at the menopause was followed by measuring gonadotrophin to creatinine ratios in small samples of urine. For follicle stimulating hormone, values of the ratio corresponded closely with both plasma concentration (correlation coefficients: 0.93, 0.94 and 0.93 for first morning, mid-morning and mid-afternoon urine samples: all results expressed as logarithms) and 24-h urine output (0.98, 0.96 and 0.94). Comparable figures for luteinising hormone (LH) were 0.94, 0.94 and 0.90; and 0.95, 0.95 and 0.90. Multiplication of the gonadotrophin to creatinine ratios (I.U./mmol) by 10 gave a figure close to 24-h gonadotrophin output (I.U.).


Immobilized enzyme nylon tube reactors: creatininase and creatine kinase linked to the pyruvate kinase-lactate dehydrogenase indicator system in the detection of creatinine and creatine.

A four enzyme system catalyzing a sequence of reactions was immobilized onto the inside surfaces of nylon tubes and studied as a possible method of analysis for the first substrate, creatinine in the sequence of reactions. The system was first studied in solution as a guide to optimizing conditions for preparation and use of the immobilized enzyme nylon tube reactors. The scope and limitations of this approach to routine analysis is discussed.


Immobilized Enzyme Pipette, Impette. Routine estimation of serum and urine urea.

This paper describes the preparation, properties, principles of operation and use of a new device, Immobilized Enzyme Pipette or Impette in routine determination of blood and urine urea. Clinical trials carried out in a medium sized hospital gave very reliable and reproducible results with a very high precision.


Abnormalities of erythrocyte membranes in biliary atresia: ultrastructure and lipid composition.

Biochemical and ultrastructural studies of red blood cell membranes from seven patients with congenital biliary atresia have been performed. Scanning electron microscopic observations revealed that more than half of these cells were of abnormal shapes, such as target, spur and cup-formed cells. By freeze-fracture electron microscopy, membrane particle-free smooth areas were noted in the fracture faces. In addition, the number of membrane-associated particles was 20% less than those in control subjects. The lipid analysis of red blood cells showed a significant increase in phospholipid and a marked increase in cholesterol content. The increase of phospholipid content was primarily caused by the increase of lecithin. The acyl chain analysis of erythrocyte lecithin demonstrated an increase in palmitic, palmitoleic and oleic acid, and a decrease in stearic and linoleic acid. These observations are similar to those of acquired biliary obstruction. The fluidity of erythrocyte membrane lipid, studied by a fluorescence technique using fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH), was found to be less in an individual with congenital biliary atresia than in the control subject.


Orthostatic postural activity disorders recorded by statokinesimeter in post-concussional syndromes: oculomoter aspect.

The purpose of this paper is to emphasize the influence of the mesencephalic oculomotor system regulation on orthostatic postural regulation. The body at rest is never immobile but it swings continuosly. The amplitudes and the frequencies of body sway give an indication of the function of the different sensorimotor loops which are involved in body balance. An apparatus, the statokinesimeter, allows us to record these motions by means of displacement of the body's centre of gravity with respect to the centre of the basis. In the case of head or cervical injuries, the brain stem is often implicated giving a partial nuclear unilateral lesion of the III nucleus; double vision of a few degrees with picture separation of never more than 4 degrees occurs. A compensory deviation of the head follows. A conflict between afferents from the III, IV, VI, VII, XI and supraspinal nuclei occurs. The amplitude and the frequency of the body sway change. A form of treatment is proposed for the 'pseudo-vertigo' of post-concussional syndrome.


Lateral cervical anomaliew and salivary heterotopia.

It is widely accepted that most lateral cervical fistulas with an external opening in the suprasternal region orginate from second branchial arch remnants. Lateral cervical cysts occur higher in the neck and according to current theories, may develop either from the branchial apparatus or from glandular inclusions in cervical lymph nodes. An histological study revealed salivary tissue in 9 of 24 fistulas, 5 of these representing a congenital cervical salivary sinus, while only one of 17 cysts showed salivary heterotopia. This investigation indicates that salivary tissue may occur more commonly in the lateral neck than is generally realised, and that lateral cervical cystis and fistulas probably differ in their histogenesis.


Bell's palsy; prognostic accuracy of case history, sialometry and taste impairment.

A prognostic test in cases of Bell's palsy must be accurate and applicable early in the course of the disease. The case history is examined in 239 patients with Bell's palsy. The prognostic value of the case history, taste impairment and sialometry is investigated in a series of untreated patients who were followed up for 12 months. Sialometry is advocated as a reliable progostic indicator in the individual patient if a strong stimulant is used. None of the other examined parameters discrimates between good and poor outcome with reasonable accuracy. A follow-up time of 12 months is advocated in determining the ultimate outcome of a Bell's palsy.


Physiological changes in the treatment of acrophobia (fear of height).

In order to investigate the physiological changes produced by the treatment of acrophobic patients body movement and Microvibration were measured before and after treatment. Eighteen acrophobic patients were assigned at random to 1 of the 2 groups: a treatment group (n = 8) and a non-treatment group (n = 10). The control group consisted of 16 healthy volunteers. Body movement area while viewing a slide of a high place or imagining a high place in the treatment group decreased significantly after treatment. Body movement of the control group showed almost no changes, and that of the non-treatment group was situated between the above-mentioned 2 groups. Simple body movements of the acrophobic patients without any stimulation of height were bigger than those of the control subjects. MV pattern of the treatment group had a tendency to improve under psycho-therapy. Acrophobic patients had more abnormal MV patterns than the normal subjects.


BIPP induced artefacts on computerized tomography.

A case of BIPP induced artefacts on computerized tomography (CT) is presented. Because of the value of CT as a non-invasive intra-cranial investigation, the use of radio-opaque materials in the site to be investigated is to be avoided. In cases likely to require the use of CT, a substitute for BIPP packs in ear surgery is now required.


Lyon phenomenon in ouabain-treated erythrocytes of Duchenne muscular dystrophy carriers as revealed by cell electrophoresis.

Using the cell-electrophoretic method, 0.1 mM ouabain/l Eagle MEM medium caused a moderate decrease in electrophoretic migration time (EMT) of erythrocytes in 19 out of 23 DMD-affected patients. In 8 out of 10 healthy boys, 0.1 mM ouabain induced an increase in erythrocyte EMT. Under the influence of 0.1 mM ouabain, the erythrocytes clearly showed a bimodal distribution of EMT in 9 out of 13 definite carriers and in 8 out of 17 possible carriers. In DMD patients, and in 10 healthy boys and 10 healthy women, no bimodal distribution of EMT was observed.


A pericentric inversion of chromosome 9 and a rearrangement involving chromosomes 9 and 10, observed in two generations. Clinical description of chromosome 9 (p12-p21) deletion syndrome.

The clinical picture associated with a deletion of a central part of the short arm of chromosome no. 9 is described in two siblings. The clinical signs differ from those described in deletion of the terminal part of the short arm. Pericentric inversion of chromosome no. 9, combined with a rearrangement involving chromosomes 9 and 10, was found in the mother and the maternal grandmother of the propositus.


High alkaline phosphatase activity in isoproterenol stimulated fibroblast cultures from patients with numerically unbalanced chromosomal aberrations.

Treatment of cultured fibroblasts from patients with unbalanced chromosomal aberrations with a mixture of isoproterenol, theophylline and ascorbic acid resulted after 48 hours in an at least three-fold increase of alkaline phosphatase activity on a per cell basis, whereas cells from normal healthy individuals did not show this dramatic response. Cells were studied from patients with trisomy 21 (14 cases), trisomy 18 (3 cases), trisomy 13 (1 case), pentasomy X (1 case), Turner syndrome (2 cases), and Klinefelter syndrome (1 case), and no exception was noted. The mechanism of this phenomenon is not clear, but it is speculated that increased cyclic-AMP levels caused by the action of isoproterenol on adenylcyclase may account for excessive reactions of unbalanced cells as compared to normal cells. This simple biochemical diagnostic procedure might become useful in screening programs for unbalanced chromosomal abberations.


Prenatal diagnosis of thrombocytopenia with absent radii.

Six pregnancies at risk for thrombocytopenia and absent radii (TAR) have been studied between 16 and 20 menstrual weeks utilizing fetal radiography. Two affected and four unaffected fetuses have been correctly identified. Fetal radiography can provide reliable prenatal diagnosis in pregnancies at risk for TAR.


Chromosomes in retinoblastoma patients.

In a series of eight patients with retinoblastoma, one was found to have a reciprocal translocation of chromosomes 1 and 13. The breakpoint on chromosome 13 is at band q12, which suggests that the retinoblastoma locus is less distal than previously thought.


Additions to the myotonic dystrophy linkage group.

One hundred and thirty members of a family with 33 cases of Dystrophia myotonica (Dm) were tested for various genetic markers including Km, Jk, Lu and Se. All individuals tested were Lua negative, but many were heterozygous for the other markers. Except for two cases all patients with Dm were positive for Km3 and Jka and they were secretors. Assuming the genes are linked and that the order of the loci is Km, Jk, Lu, Se, Dm, then there were 22 meioses informative for Dm and in 10 of these a recombination must have occurred between two of the five loci. Looking at individual pairs of loci, there were no recombinations between Km and Jk in seven informative meioses, three recombinations out of 10 meioses informative for Jk/Se, no recombinations between Se and Dm in five informative meioses and three recombinations out of 12 meioses informative for Jk/Dm.


Satellite DNA loss and nucleolar organiser activity in an individual with a de novo chromosome 13,14 translocation.

The distribution of satellite DNA and nucleolar organiser activity have been studied in a female with a new dicentric translocation chromosome derived from the maternal chromosomes 13 and 14. More than half the satellite DNA (60.5%) was lost in the translocation, together with both the nucleolar organiser regions (NOR'S). However, at least one NOR (chromosome 21) which was inactive in the mother (by the AgI reaction) is active in the subject, and this may be an example of functional compensation. The somatic cells of the mother of the subject, which do not have the translocation, show a high frequency of acrocentric associations, but these do not include any obvious excess of associations involving chromosomes 13 and 14, indicating that the high frequency of association in somatic cells is not in itself a predisposition to Robertsonian translocation in germ line cells. The father's chromosomes 9 both have more satellite DNA in the secondary constriction than normal, but this is not reflected in any obviously larger size of the C-band in this region.


Partial trisomy 1q syndrome.

Six cases of partial trisomy 1q, including four cases from the literature and our own two observations are summarized with respect to their clinical symptoms. Distinct similarities of the external aspect and of internal malformations allow the delineation of a syndrome of partial trisomy 1q.


Histochemical studies for 5'-nucleotidase and alpha-naphthyl (non-specific) esterase in lymphocytes from patients with primary immunoglobulin deficiencies.

Lymphocytes from patients with primary immunodeficiency were tested histochemically for ecto 5'-nucleotidase (5'N) and alpha-naphthyl (non-specific) esterase. More than half the patients with 'common variable' hypogammaglobulinaemia, all those with X-linked hypogammaglobulinaemia and some of those with selective IgA deficiency had a very low percentage of lymphocytes staining for 5'N as compared to controls. A lack of B cells probably explains the finding in X-linked hypogammaglobulinaemia, but does not fully explain the results in the other groups. Most patients with 'common variable' hypogammaglobulinaemia had a very low percentage of lymphocytes with granular staining for alpha-naphthyl (non-specific) esterase in contrast to normal numbers in those with X-linked hypogammaglobulinaemia and most of those with selective IgA deficiency. Granules containing non-specific esterase are characteristically found in 'mature' T lymphocytes. The enzyme abnormalities in the T and B cells of 'common variable' hypogammaglobulinaemic patients could be explained by 'immature' cell types.


Quantitative and qualitative investigations of serum IgG subclasses in immunodeficiency diseases.

Determinations of IgG subclasses were made by electroimmunoassay and crossed immunoelectrophoresis, and Gm markers were typed in sera from seventeen patients with well-defined immunodeficiency diseases. Certain IgG subclass and Gm patterns were recognized in various diseases: IgG2 deficiency and homozygosity of Gm (4,5) in the cartilage-hair-hypoplasia syndrome, in the ataxia telangiectasia syndrome and in selective IgG subclass deficiency; and IgG3 deficiency and homozygosity of Gm(1,-5) in the Wiskott-Aldrich syndrome. The findings suggest a common structural or regulator gene defect in some immunodeficiency diseases. In IgA deficiencies, the levels of IgG1 were raised. In patients with IgG subclass deficiencies there was sometimes a compensatory increase of the remaining IgG subclasses, with a preponderance of IgG1 and IgG3. The increased Ig1 showed restricted heterogeneity with only an increase of the electrophoretically cathodal part. This part contained both kappa and lambda chaings. IgG subclass deficiency indicates treatment with gammaglobulin even if the serum levels of IgG are normal or increased.


Complement activation in asymptomatic patients with sickle cell anaemia.

Previous reports have suggested that a defect in serum complement may contribute to the increased susceptibility to infection shown by patients with sickle cell anaemia (SCA). In order to define the nature of any complement abnormality in SCA, we investigated the complement system in eighty-seven patients during asymptomatic periods, and analysed factor B turnover in a small sample. In these patients geometric mean serum concentrations of functionally active factor B and factor D, and of C3 and C4 protein (expressed as a percentage of normal reference serum) wer lower than in controls (78% vs 107%, P less than 0.001, 86% vs 103%, P less than 0.001, 91% vs 100%, P less than 0.01, 89% vs 105%, P less than 0.05 respectively). The ratio of the serum concentration of functionally active factor B to factor B protein was lower in patients than in controls (means 75% s.d. 16% vs mean 93%, s.d. 22% P less than 0.001), indicating a functional deficiency of factor B protein. In addition, the fractional catabolic rate of radiolabelled factor B was markedly increased in four out of seven asymptomatic patients studied, and was inversely related to the functional factor B concentration in serum (r = -0.59, P less than 0.05); factor B synthesis was uniformly increased. Complement activation was not related to the presence of circulating C1q binding material. We conclude that complement activation, rather than defective synthesis as previously suggested, contributes to the abnormalities in complement componenet concentration and function in asymptomatic subjects with sickle cell anaemia.


Effect of sex hormones on the immune system of guinea-pigs and on the development of toxoplasmic lesions in non-lymphoid organs.

The role of cell-mediated immunity in the pathogenesis of toxoplasmic lesions in non-lymphoid organs and its significance in resistance to infection with Toxoplasma gondii were studied in guinea-pigs. Alteration of the sex hormonal environment in both sexes has significant effects on the immunological status of the guinea-pig, changing its reaction to toxoplasmic infection. Thus, gonadectomy appears to enhance delayed hypersensitivity, leading to a greater prominence of lesions in the non-lymphoid organs of gonadectomized male and female animals than in controls. This observation supports the possibility that delayed hypersensitivity plays a major role in the pathogenesis of non-lymphoid, toxoplasmic lesions. Hexoestrol administration for a long period of time induces complete thymic atrophy and severe involution of the thymus-dependent areas of the lymph nodes and spleen, leading to the suppression of cell-mediated immunity and overwhelming disease in those animals infected with Toxoplasma gondii, suggesting that cell-mediated immunity is of importance in resistance to toxoplasmic infection. In view of these findings, the difficulties in the treatment of non-lymphoid toxoplasmic lesions, such as ocular ones, by depressing cellular immunity, are discussed.


Antibody-dependent and phytohaemagglutinin-induced lymphocyte cytotoxicity in systemic sclerosis.

Cell-mediated cytotoxicity was examined in thirty-seven patients with systemic sclerosis using both whole blood and purified peripheral blood mononuclear cells (PBM) to measure antibody-dependent (ADCC) and phytohaemagglutinin (PHA) induced lymphocyte cytotoxicity to 51Cr-labelled Chang liver cells. In twenty-three mildly affected patients, ADCC and PHA-induced cytotoxicity did not differ from that found in control populations. By contrast, fourteen patients severely affected by extensive visceral disease showed reductions in both ADCC and PHA-induced cytotoxicity which were more marked in whole blood assays (P less than 0.001) than in those performed with PBM (P less than 0.05). The addition of patient's sera to control cytotoxicity assays suggested that blocking or suppressive serum factors could only account for some of the disproportionate reduction in whole blood cytotoxicity which, in the main, must be due to a lack of circulating effector cells. These results are in agreement with previous findings of reduced numbers of circulating thymus-dependent lymphocytes in patients with severe disease, a defect of cell-mediated immunity that may result from the chronic antigenic stimulation of an autoimmune disease process.


The quantification of viable Leishmania enriettii from infected guinea-pig tissues.

Results are presented demonstrating that in vitro cultivation of Leishmania enrietti can be used to determine the level of viability in suspensions of L. enrietti used for infection and also to quantify the number of viable organisms in infected tissues.


Immunological aspects of cryoprecipitates from the sera of chronic HBsAg carriers.

The sera of chronic hepatitis B surface antigen (HBsAg) carriers and seropositive controls were examined for the presence of immune complexes by cryoprecipitation. Cryoprecipitates (CP) were tested for HBsAg, antibody to HBsAg (anti-HBs), major classes of immunoglobulins, components of the complement system, rheumatoid factor and the ability to activate the alternative pathway of the complement system. For this analysis the methods employed included: radioimmunoassay, reverse passive haemagglutination, immunofluorescence, sucrose density gradient ultracentrifugation, agar-gel diffusion, immunoelectrophoresis, counterimmunoelectrophoresis, latex agglutination, and a haemolytic method for the detection of the activation of the alternative pathway of the complement system. HBsAg was frequently observed in the CP from chronic HBsAg carriers. No anti-HBs activity was detected in the serum of chronic HBsAg carriers. However, the CP from a number of chronic HBsAg carriers contained immunoglobulins and components of the complement system in the absence of rheumatoid factor, anti-HBs activity and were able to activate the alternative pathway of the complement system. On immunoelectrophoresis, a component of the CP reacting with anti-IgG, ANTI-IgA and anti-HBs antisera and demonstrating an altered (faster) electrophoretic mobility was observed. The nature of the CP strongly suggests the presence of circulating immune complexes in asymptomatic chronic HBaAg carriers. These immune complexes may be important in the eventual expression and outcome of clinical disease in apparently healthy carriers of HBsAg.


Temporal relationship between cardiac performance and myocardial uptake of dl-propranolol in man.

1. The temporal relationship between the myocardial arteriovenous difference of plasma propranolol concentration and heart rate and left ventricular dp/dt maximum was examined for 30 min after intravenous administration of 10mg (0.04 mmol) dl-propranolol in nine patients with ischaemic heart disease. 2. Very large positive myocardial a.v. differences of propranolol within the first 5 min became negative after 5 min, indicating a rapid, avid uptake with subsequent release of propranolol from the heart. 3. The heart rate and left ventricular dp/dt maximum increased 1 min after propranolol administration and did not change after the fifth min of the observation period. 4. The data suggest that a large number of unspecific binding sites for propranolol are present in the heart in addition to the specific beta-receptors.


Drug interactions in cat isolated tracheal smooth muscle.

1. The interactions between some drugs that contract and relax airways smooth muscle have been investigated in the cat isolated trachea. 2. Isoprenaline and theophylline inhibited serotonin-elicited contractions more than acetylcholine-mediated responses. This was observed both in terms of the degree of inhibition and the concentration of the relaxant drug producing this inhibition. 3. The acetylcholine- and serotonin-induced contractions were inhibited more by theophylline than by isoprenaline displaced acetylcholine and serotonin response curves to the right whereas theophylline caused a flattening of the curves. 4. Isoprenaline was more effective in hibiting serotonin contractions than acetylcholine contractions when the tracheas were bathed in K+ depolarizing solution, suggesting that the difference in the suscepbibility of serotonin and acetylcholine contractions to isoprenaline was not dependent on the electrical membrane potential. 5. Isoprenaline inhibited the tonic component of acetylcholine contractions more than the phasic component. 6. The differences in the pharmacological responses to the contractile and relaxant drugs in cat tracheal preparations provide further examples in smooth muscle of different mechanisms by which acetylcholine and serotonin induce contraction and isoprenaline and theophylline relaxation.


Measurement of clofibric acid (CPIB) metabolites in plasma of patients on clofibrate therapy.

1. The metabolites of clofibric acid [CPIB;2-(chlorophenoxy)-2-methylpropionic acid] are present in the plasma of patients on clofibrate therapy. The highest plasma concentrations of CPIB in metabolite form (up to 51 micrograms/ml) were generally found in patients with renal disease. Negligible concentrations (less than or equal to 2 micrograms/ml) were found in only seven patients out of thirty-six studied. 2. The two conjugates of CPIB found in urine were present in plasma. 3. When measuring conjugated CPIB in plasma it is essential to take care in the handling and storage of specimens, and to select an assay method known to be specific for unmetabolized CPIB.


Occurrence of excitatory dopaminoceptors in the rat and guinea-pig vas deferens.

1. Noradrenaline and dopamine produce contractions of isolated preparations of vasa deferentia from the rat or the guinea-pig. 2. Dopamine was three to five times as potent as noradrenaline on the rat vas deferens, but was considerably less potent than noradrenaline on the guinea-pig vas deferens. 3. Phentolamine antagonized responses to noradrenaline and dopamine of vasa deferentia of both species. On the rat vas deferens the pA2 values against noradrenaline and dopamine were 7.6 and 7.4, respectively. 4. Pimozide, a dopamine antagonist, antagonized responses to dopamine on vasa deferentia of both species, but was not significantly active in antagonizing responses to noradrenaline. On the rat vas deferens, the pA2 value against dopamine was 7.9. t. The beta-adrenoceptor blocking agents, propranolol and oxprenolol slightly enhanced dopamine-induced contractions on vasa deferentia of both species. 6. It is suggested that dopamine might act on both a-adrenoceptors and dopaminoceptors in the vas deferens, and that the component due to activation of dopaminoceptors is greater in the rat than the guinea-pig.


Liver alcohol and aldehyde dehydrogenase: inhibition and potentiation by histamine agonists and antagonists.

1. The in vitro effects of histamine, some other H1- and H2-receptor agonists and some antagonists were studied on the specific activities and kinetics of rat liver alcohol dehydrogenase (ADH), and cytoplasmic and mitochondrial liver aldehyde dehydrogenase (ALDH). 2. Histamine (H1- and H2-agonist) non-competitively inhibited ADH. There were no changes of cytoplasmic and mitochondrial liver ALDH activities in the presence of 2-(2-aminoethyl) pyridine. 3. Betazole (H2-receptor agonist) produced a competitive inhibition of mitochondrial ALDH but not of ADH or cytoplasmic ALDH. 4. Diphenhydramine (H1-receptor antagonist) non-competitively inhibited ADH at a lower concentration. It stimulated mitochondrial ALDH activity without changes in cytoplasmic ALDH from control values. 5. Burimamide (H2-receptor antagonist) produced a biphasic and dose-dependent stimulation and non-competitive inhibition of ADH and it non-competitively inhibited ALDH in both cytoplasmic and mitochondrial fractions. Metiamide (H2-receptor antagonist) non-competitively inhibited all ADH and ALDH of both liver fraction studied. 6. It is concluded that liver ADH and ALDH activity can be altered by compounds which affect both H1- and H2-histamine receptors and that these compounds may cause an in vivo potentiation and/or reduction of the toxic effect of ethanol.


The effect of delta 9-tetrahydrocannabinol (delta 9-THC) on the turnover rate of brain serotonin of the rat.

1. One isotopic and three non-isotopic methods were used to determine the effect of an acute intravenous dose of delta 9-tetrahydrocannabinol (delta 9-THC, 2 mg/kg) on the rat brain turnover rate of serotonin. 2. In control animals the turnover rate of serotonin was about 2 nmol/g per h. This rate was not altered by delta 9-THC when it was calculated from the rise of 5-hydroxyindoleacetic acid following probenecid from the rise of serotonin following pargyline. 3. delta 9-THC did not alter serotonin turnover rate when it was calculated from the conversion of 3H-tryptophan to 3H-serotonin. 4. The serotonin turnover rate was significantly increased by delta 9-THC when the rate was calculated from the decline of 5-hydroxyindoleacetic acid following pargyline. 5. These results suggest that delta 9-THC does not alter the turnover of rat brain serotonin. The previously reported delta 9-THC-induced changes in body temperature and increased brain levels of 5-hydroxyindoleacetic acid may be mediated by some other mechanism such as interference by delta 9-THC of the vesicular binding of serotonin.


Adult bacteremia caused by gram-negative bacilli.

Because gram-negative bacteremia is increasing in frequency, it is important for physicians to suspect, carefully evaluate, and initiate early appropriate therapy. Identification of the probable source of bacteremia and a knowledge of the hospital microflora are helpful in guiding antibiotic selection. The mortality of gram-negative bacteremia tends to vary with the severity of the underlying disease and, accordingly, tends to be low in obstetrical patients. Prevention is the most effective method of controlling these infections and, therefore, care and discretion when employing invasive devices is essential. With the increasing incidence of these infections and the increased threat of antibiotic plasmid resistance, future infection control in high-risk patients by immunoprophylaxis may be necessary.


Ambulatory evaluation and treatment of infertile couples.

With the exception of major surgical procedures, all investigative and therapeutic regimes regarding infertility can be instituted and carried to completion via the ambulatory office and hospital route if a foundation for understanding and cooperation between the physician and the couple is established during the initial stages.


The scintigraphic visualization of a lingual thyroid after excision of associated ectopic thyroid tissue.

The scintigraphic findings in a 2-year-old boy with both a lingual thyroid and ectopic thyroid tissue in the neck are presented. One day following excision of the neck mass, no functioning thyroid tissue was seen. However, one month later, the lingual gland was well visualized. The possible reason for this is discussed.


Assessment of left ventricular function in aortic insufficiency using echocardiography, gated scintigraphy and contrast angiography.

Echocardiography, gated scintigraphy and contrast angiography were used to measure left ventricular ejection fractions in 20 patients with varying degrees of aortic insufficiency. There was good correlation between the radionuclide and the angiographic ejection fractions. Echocardiographic ejection fraction correlated less well with the angiographic ejection fraction. The radionuclide left ventricular ejection fraction may prove valuable in the noninvasive serial evaluation of left ventricular function in patients with chronic aortic insufficiency.


Rapid redistribution of thallium-201 post stress testing in a patient with variant angina.

Reperfusion of an ischemic area after stress scanning with 201TI can occur rapidly and early scanning post stress testing is emphasized.


Radionuclide Meckelogram with particular reference to false-positive results.

Abdominal imaging 99mTc-pertechnetate is a most sensitive and noninvasive procedure for detecting Meckels diverticulum. However, false-positive results are unavoidable. We report two in which false-positive results were obtained and present a summary of reported false-positive results.


Diagnostic features of a unilateral fused kidney with a lower-pole inflamed, hemorrhagic cyst.

Numerous imaging procedures contributed to the diagnosis of unilateral fused kidney with an associated lower-pole mass in a 68-year old man with back and right lower quadrant pain. 99mTc-DMSA renal scintigraphy provided the best global picture of the renal anomaly and a lower-pole contour defect suggesting a mass.


False-positive 111In-labeled leukocyte scan in cystic fibrosis.

A false-positive 111In-labeled leukocyte scan was obtained in a girl with cystic fibrosis and infection. There was focal accumulation of 111In activity in the abdomen and marked pulmonary uptake. Surgical exploration and postmortem examination revealed no abdominal abnormality. The abnormal activity was apparently due to swallowed sputum which contained labeled leukocytes. Broader clinical implications are discussed.


99mTc-sodium pertechnetate permeation into cerebrospinal fluid.

The integrity of the blood-brain barrier may be reflected by the blood and cerebrospinal fluid (CSF) if these two compartments are sampled at an appropriate interval after the administration of certain substances. After reading through the controversial literature, this study was undertaken to determine the frequency with which 99mTc-sodium pertechnetate entered the CSF to an abnormal extent, and to see whether this was related to any particular pathology. The plasma-to-CSF ratios were determined in 51 patients 4 hours following the intravenous administration of 0.5-10 mCi 99mTc-sodium pertechnetate. In 23 patients with active CNS tuberculosis, the mean value of the pertechnetate plasma-to-CSF ration was 32.12. In contradistinction, the plasma-to-CSF pertechnetate in 28 nontuberculous subjects was considerably higher (144.63). For the purpose of correlation, a 82Br partition test was also done on each subject 48 hours following the oral administration of NH82Br. The 48-hour study in each of these cases was generally in agreement with the pertechnetate studies: the plasma-to-CSF ratio mean value was 1.25 in patients with active CNS tuberculosis, while for the nontuberculous patients it was 3.32.


Bone marrow scintigraphy in the evaluation of acute nonlymphocytic leukemia.

The clinical usefulness of bone marrow scintigraphy in evaluating adult acute nonlymphocytic leukemia (ANLL) was assessed through 39 studies of 23 adult patients. Nineteen patients were in various stages of relapse and remission of ANLL and four patients were preleukemic. Results of scintigraphic studies were correlated with information concerning bone marrow status, therapy, and subsequent clinical response. When 99mTc-sulfur colloid was utilized, eight of 34 studies were normal, but in the majority of patients there was an abnormal distribution of bone marrow reticuloendothial activity. Three of the patients with normal studies were in complete remission at the time of the normal scan. The duration of these remissions tended to be long. In four patients, 67Ga-citrate scintigraphy showed a wide variation of bone marrow uptake despite a marked increase in bone marrow myeloblasts. There was no significant correlation between scintigraphic distribution and bone marrow morphological status nor with subsequent ability to induce a complete remission with chemotherapy. Nevertheless, these studies raise the possibility that bone marrow scintigraphy may be useful in predicting the stability of an induced complete remission in ANLL. Confirmation of this possibility must await further studies.


Plasmacytoma simulating meningioma on brain scan.

The flow study and static images of the brain scans of two patients with plasmacytoma of the skull were identical to the pattern most commonly seen with meningioma. There was a prominent blush on dynamic imaging which did not wash out combined with an intense uptake on static views adjacent to a meningeal surface. In patients with known multiple myeloma, the diagnosis of plasmacytoma of the skull or cranial cavity should be considered when this characteristic pattern is seen.


Radiophosphate imaging of regional migratory osteoporosis.

A patient with regional migratory osteoporosis had somewhat unusual features in that there was involvement of the small joints of the foot demonstrable by 99mTc-methylene diphosphonate joint imaging.


Usefulness of 99mTc-methylene diphosphonate scintigraphy in nephrocalcinosis.

Abnormally increased uptake of 99mTc-methylene diphosphonate in the kidney parenchyma was found in a patient with acutely exacerbated sarcoidosis. The findings could be related to active sites of nephrocalcinosis.


Radionuclide computerized ventilation studies.

Studies of regional lung function have developed rapidly in the last few years. Xenon -133 is the most frequently used isotope because of its physiocochemical and biologic properties. Fixed counters are being replaced by the scintillation camera interfaced to a computer allowing easy acquisition and interpretation of results and numerical data. The use of isotopes other than 133Xe, such as 81mKr, is less widespread. In this article, a method is described for studying regional ventilation in which the isotope used is 133Xe, the radioactivity is recorded by a scintillation camera, interfaced to a computer andsix (or 12) regions for both lungs are selected and investigated. The regional ventilation is expressed as an absolute value in ml/min/ml of ventilated volume as well as the relative contribution in percent of each zone to the total ventilation and volume. To illustrate this method, the results are reported for healthy subjects and for patients with chronic bronchitis and emphysema. Scintigraphs of individual cases are presented. Certain theoretic problems related to calculation of regional ventilation are discussed.


Behavior modification therapy in hyperactive children. Research and clinical implications.

One hundred fifty-seven studies employing behavior modification in the management of hyperactive and disruptive children were reviewed. The studies were analyzed against standards of scientific validity. The review found: (1) behavior modification was effective in alleviating problem behaviors; (2) token programs were the most commonly used; (3) both positive reinforcement and punishment were effective; positive reinforcement, however, had the advantage of improving self-esteem; (4) behavioral problems occurring in the home most likely require a home-based program; (5) behavior modification and stimulant medication can be used simultaneously, often with additive effects; and (6) long-term benefits beyond one year have not been assessed.


Forecasting individual pharmacokinetics.

Often drug dosage may be chosen rationally by use of plasma concentration (CP) as the "therapeutic" end point. The ability to accurately forecast the CP resulting from a dosage regimen is central to choosing that regimen. Tradionally forecasting has been attempted only by accounting for known influences on pharmacokinetics, such as sex, age, and renal disease. One must also adjust for previously observed CPs. Herein, we discuss and explain an approach to both of these tasks, mainly focusing on the latter. The approach balances observed outcomes against prior expectations taking account of observation CP error. For digoxin, use of 1 measured CP, as opposed to none, improves forecast precision for future CPs by 40% (decrement in variance of forecast error), and 2 CPs improve it by 67%. There is also an increase in forecast accuracy (decrement in mean of forecast error) as the number of CPs used increases. After only 2, forecast accuracy and precision are as good as theoretically possible. Moreover, information from CPs is far more valuable for forecasting than that from observable patient features-sex, age, and the like; use of all the latter information does not improve accuracy and precision as much as only 1 CP.


Plasma levels and renal excretion of phenytoin and its metabolites in patients with renal failure.

Phenytoin (DPH) and its two major metabolites, conjugated and unconjugated 5-(4-hydroxyphenyl)-5-phenylhydantoin (4-OH-DPH), have been studied in plasma and urine to 4 healthy subjects and 3 uremic patients during two weeks on DPH, 0.1 gm daily. Only 0.4% to 1.2% of the dose was excreted as unchanged DPH. The DPH concentrations in urine were in the same range as calculated unbound levels of DPH in plasma in the normal subjects; 1% to 2% of the dose was excreted as unconjugated 4-OH-DPH in the normal subjects. In the uremic patients, renal clearance of this metabolite was reduced to one-sixth that percentage. Plasma concentrations rose to values twice as high as normal, indicating increased rate of glucuronidation. Urinary recovery of conjugated 4-OH-DPH in healthy subjects was 52% to 94%. Its renal clearance was close to glomerular filtration rate when corrected for protein binding, suggesting elimination by glomerular filtration rate when corrected for protein binding, suggesting elimination by glomerular filtration only. Plasma concentrations of conjugated 4-OH-DPH reached plateau levels around day 4 in normal subjects. In the uremic patients, plasma concentrations of this metabolite accumulated to levels 10 times normal, and after 15 days of medication plateau levels did not seem to have been reached.


Influence of pentobarbital on metoprolol plasma levels.

Induction of microsomal enzymes with barbiturates in rats has little effect on the metabolism of metoprolol, compared with propranolol and alprenolol, which undergo extensive hepatic extraction in animals and man. Our study was designed to examine whether the metabolism of metoprolol is inducable by barbiturate in man. In 8 healthy subjects the area under the plasma concentration/time curve after 0.1 gm metoprolol was reduced by a mean of 32% after treatment with 0.1 gm pentobarbital at bedtime for 10 days. There was considerable interindividual variability in the reduction after pentobarbital treatment (2% to 46%).


Hemodynamic effects of intravenous timolol in coronary artery disease.

The hemodynamic effects of intravenous timolol were evaluated in 20 patients with coronary artery disease during diagnostic cardiac catheterization. The threshold dose of 0.25 mg reduced heart rate and cardiac index by 15% (p less than 0.05), left ventricular work index by 21% (p less than 0.05), and left ventricular dp/dt by 16% (p less than 0.05) while increasing left ventricular end-diastolic pressure by 49% (p less than 0.01), mean pulmonary arterial pressure by 17% (p less than 0.01), and systemic vascular resistance by 16% (NS). Larger doses (0.5 mg and 1.0 mg) induced similar responses with a greater effect on systemic vascular resistance (+22%, p less than 0.01, and +31%, p less than 0.001). The mean arterial pressure and stroke volumes were not affected by timolol. Peak effects, occurring at about 10 min after drug injection, did not correlate with plasma levels. The overall hemodynamic effects of timolol were similar to those reported for equipotent doses of propranolol and could be accounted for by the beta-adrenoceptor blocking activity.


Morphine kinetics in children.

The kinetics of morphine in the plasma of children (0 to 15 yr) can in most cases be adequately described by a triexponential function. When the dosage in children receiving morphine as premedication before surgery is based on kilograms of body weight, there are only minor differences in the kinetic patterns of morphine at different ages (0 to 1, 1 to 7, and 7 to 15 yr). No significant difference in the morphine kinetics was observed between patients (7 to 15 yr) receiving the drug only as premedication and those who also received doses during surgery. The minimum morphine concentration in plasma necessary to suppress the clinical signs of pain during surgery was found to be 65 ng/ml (95% confidence limits of 46 to 83 ng/ml). Based on these minimum effective plasma levels of morphine, there does not seem to be any difference between children of different ages in their sensitivity to morphine. No difference was found in the minimum effective level of morphine when clinical signs of pain were observed by different anesthesiologists.


Kinetics and metabolism of carbamazepine during combined antiepileptic drug therapy.

The kinetics of carbamazepine using 15N-carbamazepine were investigated in epileptic patients during combined anticonvulsant therapy. The 15N-carbamazepine plasma half-lives ranged from 5.0 to 13.6 hr with a mean of 8.2 hr. These half-lives are appreciably shorter than reported during chronic carbamazepine monotherapy. Predicted steady-state plasma levels and observed plasma levels of carbamazepine were in excellent agreement. Between 32% and 61% of the dose administered is excreted in the urine as carbamazepine-trans-diol, 5.2% to 8.8% as 9-hydroxymethyl-10-carbamoyl acridane, 1% to 1.4% as 10,-11-carbamazepine epoxide, and 0.5% as carbamazepine. The data indicate that it is the epoxide-diol pathway which is induced during long-term treatment. Concomitant therapy with primidone, phenytoin, phenobarbital, ethosuximide, or methsuximide further induces carbamazepine metabolism.


Salicylate clearance, the resultant of protein binding and metabolism.

Steady-state plasma salicylate concentrations and protein binding were examined in 9 normal subjects to determine relationships among daily dose, total and unbound salicylate concentrations, and total and unbound clearances. Aspirin doses ranging from 0.66 to 4.0 mg/kg/hr were given to steady state. Free and total salicylate concentrations were measured with spectrophotometric, fluorimetric, and equilibrium dialysis techniques. Although unbound clearance decreased over the therapeutic range, total clearance was unchanged. The former is a consequence of saturable metabolism; the latter, of saturable plasma protein binding as well as saturable metabolism. The fraction unbound increased linearly with unbound concentration. Clearance determined at 1.8 mg/kg/hr was used to predict levels obtained at higher aspirin doses. Analysis of residuals was used to ascertain the accuracy of the prediction. The coefficient of variation from prediction among subjects was found to be +/- 14%. It is concluded that, in normal subjects, salicylate clearance changes relatively little over the therapeutic range because the increasing fraction unbound compensates for decreasing clearance of unbound drug.


Effect of thiazides on serum calcium.

A retrospective study of 22 hypertensive patients who were treated with thiazide diuretics for 2 to 12 yr revealed that 36% developed transient, self-limited asymptomatic elevations of serum calcium which occurred at varying periods of therapy and returned to normal within 2 to 4 wk despite continued administration of thiazides. These episodes of hypercalcemia correlated positively with increases in total protein, albumin, and globulin. The same phenomenon of intermittent hypercalcemia occurred in a prospective study of 11 patients but not in control subjects. The mean serum total calcium of the prospectively studied hydrochlorothiazide-treated patients was found to be higher than the nonthiazide control group. This difference was due to increased protein-bound calcium. The total proteins, serum albumin, and serum beta globulins of the treated group were higher, probably due to depletion of extracellular fluid. The presence of slightly elevated serum calcium in a patient treated with thiazides appears to be a common phenomenon and, unless it is marked, should not necessarily be construed as indicating covert hyperparathyroidism.


Diazepam prevents some adverse effects of succinylcholine.

Neuromuscular, circulatory, and adverse effects of intravenous succinylcholine (SCh), mg/kg, were compared in 3 groups of 40 patients each. Group I served as control; group II received diazepam, 0.05 mg/kg, 5 min before SCh; and group III was given d-tubocurarine (d-Tc), 0.05 mg/kg, for pretreatment. Diazepam pretreatment prevented muscle fasciculations, increases in serum potassium (K+) and creatinine phosphokinase (CPK) levels, increased heart rate and arterial pressure, and postoperative myalgia associated with SCh administration. The neuromuscular blocking action of SCh was not affected. Pretreatment with d-Tc did not abolish increases in serum K+ and CPK levels, was associated with a higher incidence of postoperative myalgia, and decreased the onset and magnitude of SCh-induced muscle paralysis. Our data demonstrate that diazepam, a predominant centrally acting muscle relaxant, is more effective than d-Tc in preventing the adverse effects of SCh, a peripherally acting muscle relaxant.


Clinical trial design in cancer.

The design of clinical trials in cancer is a subject which features reasonably often among FRCR (Part 1) examination questions, and as such should be of more than passing interest to oncologists. It is also a subject which is gaining in relevance since the number of trials is increasing annually due in part to the many chemotherapeutic regimes which are being proposed. This paper which is based on a lecture given in Cambridge at the Hospital Physicists' Association Annual Conference in September 1978, is intended to act as an introduction to clinical trial design. References for further reading are given and, in particular, the extensive report on randomised clinical trials to the Medical Research Council's Leukaemia Steering Committee (Peto et al., 1977, 1978) is recommended.


The role of computed tomography in the staging of bladder cancer.

Computed tomography of the pelvis has been performed in 60 patients with epithelial bladder tumours. The CT findings have been compared with the clinical staging (T-stage), lymphography (N-stage) and wherever possible the surgical staging (P-stage). Although the intraluminal tumour was visualised in a high proportion of examinations, the greatest value of CT is in the accurate delineation of the extravesical extension of the growth. This is likely to be the primary role of CT in the staging of bladder cancer. Difficulties in detecting invasion of contiguous organs, particularly the prostate, and the failure to demonstrate nodal involvement within the pelvis were noted. The technique has clear advantages over more invasive investigations and the additional information provided over and above clinical staging is seen as a major advance in the assessment of these tumours.


Vacuum contoured, liquid crystal, dynamic breast thermoangiography as an aid to mammography in the detection of breast cancer.

New thermographic equipment is introduced in the form of liquid crystals embedded in flexible, transparent, elastomeric sheaths which may be vacuum contoured to any breast. Dynamic thermoangiography, a new physiological method of performing thermography with these liquid crystal sheaths, is presented. Thermographic changes are recorded during active skin cooling and rewarming with liquid crystals in situ. The new system eliminates skin preparation previously required. Preliminary results appear to show good correlation with conventional telethermography. Vascular, as well as thermal pathology, is accentuated with rich, high contrast, thermographically calibrated colour patterns. The new system is relatively inexpensive, simple to operate, and may be suited to a programme for the detection of breast cancer.


The chest X-ray in pulmonary capillary haemorrhage: correlation with carbon monoxide uptake.

Serial changes in carbon monoxide uptake (KCO or DLCO/VA) were used to monitor episodes of pulmonary capillary haemorrhage in Goodpasture's syndrome (13 cases), immune complex nephritis (three cases) and idiopathic haemosiderosis (one case). Air-space shadowing on the chest X-ray (on a scoring system 0 to 12) was assessed in the light of the changes of KCO. In 14 out of 27 episodes of bleeding, the rise and fall of KCO was matched in time by the appearance and disappearance of air-space shadowing on the chest X-ray. In six episodes the chest X-ray remained normal despite a rise of KCO. In two cases air-space shadowing appeared up to 48 h after the rise of KCO. On five occasions chest X-ray abnormalities preceded the rise of KCO but chest infection or fluid overload accounted for three of these. In cases with suspected pulmonary capillary haemorrhage, measurements of carbon monoxide uptake will provide additional information and will assist in the interpretation of the chest X-ray.


The chest X-ray in antiglomerular basement membrane antibody disease (Goodpasture's syndrome).

The chest radiographs of 25 patients with proven antiglomerular basement membrane antibody disease (Goodpasture's syndrome) were analysed. All except two of the patients had pulmonary haemorrhage at some stage of their disease. Altogether there were 39 episodes of pulmonary haemorrhage, 25 being relapses. During seven episodes the chest radiograph was normal. Relapses of pulmonary haemorrhage never occurred in isolation but were usually associated with infection (not necessarily a chest infection) or occasionally fluid overload. Conversely fluid overload or infection were always associated with pulmonary haemorrhage provided there were high or rising titres of circulating antibodies at the time. Therefore in a patient with antiglomerular basement membrane antibody disease, the presence of shadowing in the lung fields on the chest radiograph almost invariably means the patient has pulmonary haemorrhage whether or not pulmonary oedema or a chest infection are present. Limitation of shadowing by a fissure, loss of major portions of the diaphragmatic or cardiac silhouette, involvement of the lung apex or costophrenic angles suggest an underlying chest infection. Septal lines suggest fluid overload. Pleural effusions are seen with chest infections and fluid overload. The carbon monoxide uptake (KCO) was invariably high in the presence of pulmonary haemorrhage even if the chest radiograph was normal. A combined use of KCO and chest radiographs is the best method of monitoring lung disease in these patients.


The computerised tomographic findings in paediatric orbital tumours.

Forty-two patients with histologically proven orbital tumours are presented. CT is of value in the pretreatment assessment and in delineating the extent of the orbital tumour. The surrounding bony destruction and intracranial extension are easily assessed. Post-treatment follow-up can be closely monitored using CT.


The role of endoscopic retrograde cholangiography in the diagnosis and management of patients with primary sclerosing cholangitis.

The clinical and radiological findings in four patients with primary sclerosing cholangitis one of whom had coexistent cholangiocarcinoma, are reported. The need for surgical exploration to make the diagnosis was averted by the use of endoscopic retrograde cholangiography in one patient who was managed initially with medical treatment alone. Endoscopic cholangiography may be used to monitor the progress of the sclerosing lesions; but failure to fill the intrahepatic ducts is associated with a poor prognosis due either to the severity of the sclerosing process or the presence of coexistent cholangiocarcinoma.


Diagnostic value of percutaneous transhepatic cholangiography judged by personal experience of 58 patients.

During the last seven years percutaneous transhepatic cholangiography (PTC) using a fine needle was performed in 58 patients. The biliary tract was visualised and opacified in the 52 patients with dilated biliary ducts. The correct diagnoses were made in 51 of them. In six patients with a non-dilated biliary system, successful PTC was carried out with the correct diagnoses in four of them. There were four non-fatal complications; two septic reactions, one haemobilia, and one subcapsular haematoma, but all were controlled by conservative treatment alone. The PTC with a fine needle is an accurate, useful and safe procedure of choice for the diagnosis of obstructive jaundice and jaundice of obscure origin. It is useful in distinguishing hepatocellular jaundice from obstructive jaundice thus preventing an unnecessary operation. Failure to visualise the biliary tract by an experienced hand strongly indicates that the jaundice is non-obstructive.


Radiological findings of the urinary tract in hypospadias: a report of 110 cases.

One hundred and ten patients with hypospadias underwent excretory urography and micturating cysto-urethrography before surgical correction. Urographic studies revealed abnormalities in 50 patients (45%), mainly anomalies of ascent and rotation of the kidneys. None of these patients required corrective surgery. Voiding cysto-urethrography disclosed disease of the urinary tract in 65 of 110 patients (59%). Meatal stenosis was found in 38 patients and a rudimentary vagina or a prostatic utricle was revealed in 11 others. Vesico-ureteral reflux was present in 14 patients, seven of whom had meatal stenosis as well. Cystitis was found in eight and a stricture of the membraneous urethra in one patient. The voiding cysto-urethrogram is able to demonstrate functionally significant meatal stenosis, genital abnormalities and vesico-ureteral reflux. It can also serve as a useful baseline for comparison with postoperative studies. We therefore consider that it should be included in the preoperative work-up of every patient with hypospadias. Excretory urography, however, is perhaps indicated only in those with urinary tract infection, or in patients otherwise symptomatic.


The radiological assessment of body fat.

A radiological method for assessment of obesity is described. A radiograph of the soft tissues of the upper arm is obtained and the subcutaneous fat layer measured. The results obtained by this method have been compared with those using a skinfold caliper and with the ponderal index, calculated from the height and weight of the subject. A highly significant correlation was found. The radiological method has several advantages: it leaves a permanent record, it is accurate in obese persons when the skinfold caliper cannot be used, and unlike the ponderal index it is not influenced by weight increases due to causes other than fat accumulation.