An accurate determination of human growth hormone content in different pituitary extracts, using a radioimmunoassay with polyacrylamide gel electrophoresis as a bound-free separation system.

Human growth hormone was extrated and purified according to the method of Roos et al. (Roos, P., Fevold, H.R. and Gemzell, C.A. (1963) Biochim. Biophys. Acta 74, 525). A first control of its purification and integrity was performed through molecular weight determination by gel filtration on Sephadex G-100 and on polyacrylamide gel electrophoresis (PAGE). Its biological activity was confirmed by the growth promoted in non-hypophysectomized rats at plateau. The main object, however, was the setting up of accurate, reproducible method tha could furnish the more absolute and comparable values of radioimmunoassayable HGH content in perfect agreement with the results obtained by other laboratories. This was accomplished through a radioimmunoassay system that uses HGH labelled with 125I, where separation of the bound from the free antigen is achieved on polyacrylamide gel electrophoresis, by a modification introduced in the original method of Davis. The resulting values, extremely close to that stated by the KABI-Laboratories (Stockolm), though obtained in quite different conditions of incubation, antibody concentration and with no use of second antibody, represent a confident approach to a comparable measure of this hormone in extracts, which can also be applied to plasma determinations.


Automated urinary steroid profiles by capillary column gas-liquid chromatography and a computing integrator.

13 Urinary steroid metabolites have been determined by automated gas-liquid chromatography with a 20-m glass capillary column and a computing integrator. Concentrations up to 2 mg/24 h computed by the integrator compare well with those obtained by peak height measurements. At higher concentrations discrepancies occurred, paticularly for the C21 steroids where falsely low values were calculated using peak heights. Mean excretion by healthy males and females of seven steroid metabolities is presented.


A simple radiochemical assay of thromboxane B2, 12-hydroxyeicosatetraenoic acid (HETE) and 12-hydroxyheptadecatrienoic acid (HHT) synthetized by human platelets.

A simple radiochemical assay of thromboxane B2, 12-hydroxyeicosatetraenoic acid (HETE) and 12-hydroxyheptadecatrienoic acid (HHT) produced by platelets from arachidonic acid has been described. Moreover, this assay is suitable for measuring thromboxane A2 trapped as 11-mono-O-methylthromboxane B2 and minor products such as prostaglandins E2, F2alpha and D2. The advantage of this technique is discussed as to its ease and specificity.


Serum-induced changes in electrophoretic mobility of creatine phosphokinase isoenzymes?

BB isoenzyme of creatine phosphokinase (CK-BB) obtained from human brain-extract changes its electrophoretic mobility after incubation in human serum at 37 degrees C. No change of electrophoretic mobility of CK-BB is observed after incubation in isotonic saline. We have shown by means of immunoprecipitation with specific antibodies that the structure of CK-BB is not changed. These findings are supported by other authors and make the diagnostic value of electrophoretic separation of CK isoenzymes doubtful as after a 3-h incubation CK-BB migrates similarly to CK-BB and consequently may be misinterpreted.


Serum pyruvate kinase in carriers of Duochenne muscular dystrophy.

Independent studies by two different groups (Madras and Edinburgh) have failed to confirm the suggestion that measurement of the serum level of pyruvate kinase (EC 2.7.1.40, PK) may be superior to measurement of the serum level of creatine kinase (EC 2.7.3.2, CK) for detecting female carriers of X-linked Duchenne muscular dystrophy (DMD). At present the serum level of creatine kinase remains the best test for this purpose.


Renin concentration after prolonged cold storage of human plasma.

The plasma renin concentration (PRC3.3) measured biologically or radioimmunologically after acid treatment of plasma at pH 3.3, did not change significantly during storage of human plasma at -20 degrees C for 1 year. This is completely in contradiction with the increase in plasma renin activity reported by some authors.


Free and total 3-methoxy-4-hydroxyphenylethylene glycol in human cerebrospinal fluid: relationship to blood pressure in hypertensives and patients with neuropsychiatric disease.

The concentrations of free and total 3-methoxy-4-hydroxyphenylethylene glycol (MHPG) in the cerebrospinal fluid of 43 patients were measured by selected ion monitoring gas chromatography-mass spectrometry using MHPG specifically labelled with deuterium as internal standard. Unpaired Wilcoxon comparisons of free and total MHPG concentrations in males and females showed no significant difference between the sexes. The relationships between free, conjugated and total MHPG concentrations and age, blood pressure, glucose and protein levels in cerebrospinal fluid were examined by linear regression analysis. The only significant correlations found were between free MHPG and glucose and between total MHPG and glucose.


The action of human pancreatic and salivary isoamylases on starch and glycogen.

The hydrolytic properties of the isoenzymes of human pancreatic and salivary alpha-amylase (1,4-glucan 4-glucanohydrolase, EC 3.2.1.1) were studied. The eight pancreatic isoenzymes split glycogen and starch into glucose, maltose, maltotriose, maltotetraose and oligosaccharides of 5-10 glucose units. Maltotetraose is further digested to lower homologues. The percentage of conversion to those products is dependent on the substrate and varies from one isoenzyme to another. The six salivary isoenzymes split glycogen and starch into maltose, maltotriose, maltotetraose, pannose and oligosaccharides of 5--10 glucose units. Maltotetraose and pannose are further digested to lower homologues. The percentage of conversion to these products is dependent on the substrate and is specific for each isoenzyme.


Polyacrylamide gel electrophoresis of human erythrocyte membrane enzymes solubilized with triton X-100.

Human erythrocyte ghosts were solubilized in a low ionic strength medium containing 1% Triton X-100 and subjected to electrophoresis in polyacrylamide gels containing Triton X-100. Five major bands were stained with Coomassie Blue, all except one band being heterogenous when re-electrophoresed in gels containing sodium dodecyl sulphate. It was possible to detect acetylcholinesterase, non-specific esterase, ATPase, alkaline phosphatase, 5'-nucleotidase, glyceraldehyde-3-phosphate dehydrogenase, lactate dehydrogenase, and aldolase activities on the Triton-containing polyacrylamide gels. Two of the enzymes, ATPase and 5'-nucleotidase, showed substantial inhibition by Triton X-100 in quantitative studies. This appears to be a useful method for studying membrane enzymes in normal and pathological red cells.


The evaluation of a radioimmunoassay for diphenylhydantoin using an iodinated tracer.

Radioimmunoassays (RIA) for diphenylhydantoin (DPH) are described using a rabbit antiserum to the immunogen, diphenylhydantoin-3-omega-valerate bovine serum albumin, and DPH labelled with carbon-14 (14C), tritium (3H) or iodine-125 (125I). The [125I]DPH label is prepared by iodination of DPH-3-N-ethyl-acetamido-tyrosine ethyl ester, and is stable for several months. The radioimmunoassays are rapid, precise, specific and sensitive requiring only 50 microliter serum for duplicate analyses. Serum DPH levels measured by the three RIA methods correlate well with each other (3H vs. 14C, r = 0.96 : 3H vs. 125I, r = 0.97) and with values obtained by a conventional gas chromatographic method (125I RIA vs. GC, r = 0.95). However, RIA employing the iodinated DPH tracer has significant practical advantages over the other methods and is well suited to large scale clinical application.


A new enzymatic method for determination of serum choline-containing phospholipids.

A new enzymatic method is presented for the determination of serum choline-containing phospholipids with a combined enzymatic method using phospholipase D (from Streptomyces species), choline oxidase (from Arthrobacter species) and peroxidase. The method is reproducible, and the results correlate well with those obtained by the conventional digestion method (Hoeflmayr, J. and Fried, R. (1966) Med. Ernaehr. 7, 9-10). The method affords better specificity, requires a smaller quantity of the sample and shorter time than those previously reported, and has excellent precision.


Determination of urinary estriol by radioimmunoassay. Modification of a commercial kit for serum analyses.

A simple and rapid method for urinary estriol determination is described. Modification of a commercially available kit for serum estriol gives a method for urinary estriol determination with a capacity of approximately 75 determinations a day. A high degree of accurary was revealed by indirect comparison with a mass fragmentographic method.


An improved processing of radioimmunoassay data by means of a desk-top calculator. (1) Comparison of regression procedures applied to selected kinds of radioimmunoassay.

By means of a programmable desk-top calculator, goodness-of-fit of 3 regression models, four-parameter logistic, quadratic logit-log and linear logit-log models, were evaluated by analysis of variance (F test) for data of 6 kinds of radioimmunoassays (RIA); thyroid stimulating hormone (TSH), luteinizing hormone (LH), follicle-stimulating hormone (FSH), insulin (IRI), cortisol, triiodothyronine (T3). Scatchard plot analyses were made with the representative data of these RIAs in order to find the best choice of regression model in relation to the characteristics of antigen-antibody reaction. The analysis of goodness-of-fit of the regression models by means of an F test disclosed the relation between the choice of regression models and the kinds of RIA, which could be grouped into 3 types: (1) almost identical degree of fit with any of 3 regression models (FSH and T3), (2) more or less equal degree of satisfactory fit with the logistic and quadratic logit-log models (TSH and cortisol), (3) best degree of fit with the aquadratic logit-log model among 3 (LH and IRI). The analysis of data with Scatchard plot discriminated 3 general types of curves; (1) linear (FSH) and T3), (2) linear with tail (TSH and cortisol) and (3) hyperbola (LH and IRI). From these findings, the following tentative conclusions were reached: RIA with linear pattern on Scatchard plot can be satisfactorily regressed with either of 3 models, RIA with linear with tail pattern regressed with either the logistic or quadratic logit-log model, and RIA with hyperbolic pattern regressed best with the quadratic logit-log model.


Hypocholesterolaemia in the rat after p-chlorophenoxyisobutyric acid (CPIB) administration as a synthetic triglyceride, 1,3-dipalmitoyl 2(p-chlorophenoxyisobutyroyl)-rac-glycerol.

A synthetic triglyceride analogue, 1,3-dipalmitoyl 2(p-chlorophenoxyisobutyroyl)-rac-glycerol, was fed to rats and its effect compared to that of clofibrate. Both compounds significantly lowered plasma cholesterol concentration and the decreases essentially paralleled each other, even when the dose of drug or the period of administration was varied. No consistent effect was noted on the plasma triglyceride concentration. The results suggest that synthetic lipids are potentially suitable carriers for the oral administration of pharmacologically active agents.


Serum gastrin determination with a radioimmunosorbent technique.

A low utilization of antiserum has been reported in gastrin radioimmunoassay with a solid-phase procedure. As solid-phase immunoassays possess several advantages over their soluble counterparts for the assay of many hormones, a solid-phase radioimmunoassay for gastrin was examined. The results showed that this technique was suitable for gastrin determination without being wasteful to antibodies. Sensitivity and precision of the method were comparable to other investigations on gastrin immunoassay, as well as serum gastrin levels obtained for persons with no known gastro-intestinal disease, in patients with pernicious anemia and in patients with Zollinger-Ellison syndrome.


L-Ornithine-ketoacid-transaminase deficiency in cultured fibroblasts of a patient with hyperornithinaemia and gyrate atrophy of the choroid and retina.

L-Ornithine-ketoacid-transaminase deficiency was established in cultured fibroblasts obtained from a patient with hyperornithinaemia (mean ornithine level in serum approximately 100 mumol/l) and gyrate atrophy of the choroid and retina. The deficiency was found both the L-ornithine concentrations of 3.0 mM (about twice the KM value) and 12 mM, indicating that the enzymic defect was not due to a decreased affinity for this substrate. The reliability of the colorimetric assay of ornithine-ketoacid-transaminase activity was established radiochemically. Performance of the radiochemical assay revealed the presence of an impurity in the substrate DL-[2-14C]ornithine - HCl being a strong inhibitor of the enzyme. The passage level and the subcultivation time of the fibroblasts did not influence the enzymic activity.


5'-AMP aminohydrolase activity in erythrocytes from normal and dystrophic individuals.

5'-AMP aminohydrolase activities in red blood cells from normal human adults and from patients with pseudohypertrophic dystrophy, myotonic dystrophy, limb girdle dystrophy, neuromuscular atrophy, Charcot-Marie Tooth Disease, and spinal muscular atrophy were examined. In contrast to the greatly decreased skeletal muscular levels of 5'-AMP aminohydrolase observed in Pseudohypertrophic muscular dystrophy, levels in red blood cells of all patients were not significantly different from normals. Pyruvate kinase and creatine kinase activities were determined for comparative purposes. The density distribution of normal and dystrophic erythrocytes were essentially identical.


A short, simple method for the determination of paraquat in plasma.

Paraquat was measured in plasma after precipitation of plasma proteins by addition of a solvent/salt mixture. The paraquat remaining in the supernatant is measured by its free radical formation after addition of alkaline sodium dithionite. The time required for a single determination is approximately 20 minutes.


Enzymic determination of plasma cholesterol on discrete automatic analysers.

Enzymic procedures for the determination of plasma cholesterol, using cholesterol esterase and cholesterol oxidase, have been adapted to the Vickers D-300, Vickers M,-300, and Vitatron AKES discrete analysers. The results obtained by these methods have been compared to those obtained by manual and continuous flow Liebermann-Burchard methods. The enzymic methods were found to be accurate, precise and of adequate sensitivity.


Urinary indole profile of Bufo melanostictus during hydroquinone-induced leucoderma and its regeneration.

The abnormal urinary indole profile induced by hydroquinone has been found to be reversed by psoralene, a pigmentogenic drug in vitiligo.


Microdetermination of unbound bilirubin in icteric newborn sera: an enzymatic method employing peroxidase and glucose oxidase.

An enzymatic assay method for the microdetermination of unbound bilirubin in newborn icteric sera is described. Unbound bilirubin is oxidized to colorless compounds by peroxidase in the presence of hydrogen peroxide derived from glucose by the mediation of glucose oxidase. In this method, the bilirubin is not significantly degraded before the addition of peroxidase, in contrast to the method using hydrogen peroxide. The oxidation rate is determined by spectrophotometry and chloroform extraction is eliminated. The unbound bilirubin concentration can be determined from the initial oxidation velocity of total bilirubin. The Michaelis constant, KM, was approximately 20 micrometer. The coefficient of variation for icteric serum determination was 4.4--6.5%. The concentration of unbound bilirubin was reduced after five days of storage at -20 degrees C. The bilirubin-albumin affinity was studied with purified albumin and adult serum. The dissociation constants were 2 x 10(-8) M and 5 x 10(-9) M, respectively, at bilirubin/albuminor molar ratios below 1.0. Clinically, serum samples from 75 icteric newborn infants were analysed, and the sera of premature infants were found to have remarkably high levels of unbound bilirubin compared to those of fullterm infants. The sera of a Rhesus immunization infant and an ABO incompatibility infant were remarkably higher than that of the nonhemolytic icteric sera. The unbound bilirubin concentration was also affected, in an in vitro study, by the addition of hemolysate.


[Determination of urinary alpha-keto-gamma-methylthiobutyric acid in hypermethionemia by use of gas chromatography and flame photometry (author's transl)].

In this paper we describe the determination of urinary alpha-keto-gamma-methylthiobutyric acid (ACMTB) by the use of gas chromatography of its quinoxalinol derivative and specific detection of sulphur with a flame photometric detector. Using these techniques, it was possible to study the urinary elimination of this secondary metabolite of methionine in normal children, homocystinurics and in one case of hypermethioninemia. The excretion of ACMTB is normally very weak and is strongly increased when the principal pathway of methionine is blocked.


[Heterogeneity of carbohydrate chains of acidic bronchial mucin isolated from the spatum of two subjects with chronic bronchitis].

Acidic glycoproteins having blood group H activity were isolated from the sputum of two patients suffering from chronic bronchitis by reduction of the fibrillar mucus, chromatography on ECTEOLA-cellulose and gel filtration on Sepharose 4-B. These glycoproteins were degraded with alkaline borohydride and the degradation products were fractionated by chromatography on ionexchange resins and by gel filtration. The carbohydrate chains have a wide heterogeneity with regard to acidity and molecular size. Therefore carbohydrate chain heterogeneity which was already observed for bronchial glycoproteins isolated from a patient suffering from cystic fibrosis is not specific for cystic fibrosis.


Blood and urine histamine levels in normal and pathological states as measured by a radiochemical assay.

The utility of the enzymatic radiochemical assay of histamine in diagnosing diseases with known abnormalities in histamine production was investigated. Whole blood histamine levels were abnormal only in patients with basophilia, i.e. chronic myelocytic leukemia or polycythemia vera. Histamine was not detectable (less than 1 ng/ml) in normal plasma but was detected in plasma of some patients witheither mastocytosis or chronic myelocytic leukemia. These patients also had symptoms which could be attributed to histamine release as, for example, hyperchlorhydria and hypotension. Urinary histamine excretion was also abnormally high in these diseases compared to normal subjects (range less than 5-42 microgram/24 h, n = 31). Patients with systemic mastocytosis had higher urine values (greater than 150 microgram/24 h) than those with cutaneous mastocytosis (39-88 microgram/24 h), and the urinary histamine excretion appeared to be an index of the severity of the diseases. Studies with L-histidine loading suggest that the kidney is one possible source of urinary histamine.


No inhibition by Li+ of thyroxine monodeiodination to 3,5,3'-triiodothyronine and 3,3',5'-triiodothyronine (reverse triiodothyronine).

The possibility that lithium affects the conversion of thyroxine to 3,5,3"-triiodothyronine and 3,3',5'-triiodothyronine (reverse triiodothyronine) was studied by measurement of the serum concentractions of these parameters in five patients during the first week of lithium therapy. In three patients there was a decrease in serum thyroxine concentration and a slightly less pronounced decrease in that of serum 3,5,3'-triiodothyronine. In two patients, who also received L-tryptophan or flupentixol, no change was noted in the concentrations of these compounds. There was no increase in serum 3,3',5'-triiodothyronine concentration in any of the patients. No systematic change was found in the serum concentrations of thyrotropin or unsaturated thyroid-hormone binding proteins. The results obtained do not support the contention that lithium should inhibit the monodeiodenation of thyroxine to its active and inactive metabolites.


Simplified quantitative determination of total fecal bile acids.

To determine total fecal bile acids, these are extracted with diethyl ether after boiling with a solution of potassium hydroxide in ethanediol. After evaporating the ether and dissolving the residue in methanol, the bile acids are directly determined with 3 alpha-hydroxysteroid dehydrogenase. Values for 9 normals are given.


Effect of pectin on serum cholesterol, fecal bile acids and biliary lipids in normolipidemic and hyperlipidemic individuals.

Pectin, 40-50 g/day for two weeks administered to nine normolipidemic and hyperlipidemic patients, had no effect on serum triglycerides but did cause a significant decrease in the serum total and unesterified cholesterol of hypercholesterolemic subjects in particular. This was associated with increased excretion of fecal bile acids and total steroids and increased concentration of plasma methyl sterols. Thus, the serum cholesterol reduction by pectin appears to be caused by increased cholesterol elimination into stools as bile acids which is then balanced by enhanced cholesterol synthesis. The composition of biliary bile acids and lipids was not changed and secondary bile acids and sterols decreased inconsistently in feces. The measurement of fecal dry weight suggested that the bulk of the pectin was degraded by bacteria during passage through the intestine. Consequently fecal mass and dry weight were not consistently increased, suggesting that pectin may not be an ideal fibre for increasing fecal bulk in functional colonic disorders.


Influence of glucose and inhibitors of glycolysis on release of total proteins and enzymes from human leukocytes.

We studied the influence of substrates and glycolysis inhibitors on the release of proteins and enzymes from human leukocytes. We show the relationship between metabolic level and percentage of enzyme release. Leukocyte kept alive in a nutrient-free bicarbonate medium liberate enzymes even though their metabolic state is satisfactory. The addition of glucose to themedium significantly decreases this phenomenon without affecting the energy level. Addition of glycolysis inhibitors increases it, reduces the energy level, and interferes with other metabolic pathways. The results are discussed and compared to those obtained by other authors using various cell models.


An improved procedure for the electrochemical determination of serum ionized calcium.

The "Orion model 99-20 serum calcium flow-through system" has been rendered insensitive to protein poisoning by the plascing of a dialysis membrane in front of the electrode membrane, making the addition of trypsin and triethanolamine to standards unnecessary. Flow dependency of the results obtained has been investigated.


[Determination of the IgA 7 S/IgA 11 S ratio in human bronchial secretion by thin layer gel filtration immunoelectrophoresis (author's transl)].

After extraction by treatment with Ecteola-cellulose, IgA 11 S and IgA 7 S from human bronchial secretion were first separated by thin-layer gel filtration in Ultrogel AcA 3-4, and then were analyzed by crossed immunoelectrophoresis with an immune serum against alpha chain. Precipitation areas were measured separately. A correlation factor was necessary to express IgA 7 S in terms of IgA 11 S: it was calculated with standard curves made with serum and colostral IgA.


[Exact determination of the lecithin/sphingomyelin ratio in amniotic fluid. Suggestion of a reference methods (author's transl)].

Pulmonary maturity of the fetus can be evaluated by the lecithin/sphingomyelin (L/S) ratio in amniotic fluid. To existing methods of lipid extraction, precipitation with acetone and chromatography, we add a simple and accurate estimation of sphingomyelins (S) and precipitated lecithins (Lp) without acid digestion. The method is reproducible (C.V. less than 9%) for the measurement of Lp/S ratio and gives with accuracy the concentrations of Lp, avoiding possible errors in interpretation of Lp/S. Our results show that at 35 weeks of normal gestation, Lp/S ratio is about 2 and Lp concentration, 10 mg/1.


Placental form of membrane-bound neutral arylamidase found in renal cell carcinoma.

A placental form of membrane-bound neutral arylamidase was found in the tissue of renal cell carcinoma. Membrane-bound neutral arylamidases from renal cell carcinoma, an intact part of the same kidney and placenta, had the same molecular weight (240 000) and were identical with respect to KM value and effect of inhibition by chelators or amino acids. Membrane-bound neutral arylamidase from renal carcinoma had the same electrophoretic mobility and heat stability as placental membrane-bound neutral arylamidase, but differed from kidney membrane-bound neutral arylamidase with respect to electrophoretic mobility, heat stability and susceptibility to urea inactivation. Results suggest the carcinoplacental alterations in membrane-bound neutral arylamidase isoenzyme in renal cell carcinoma.


A sensitive enzymatic method for determination of free and esterified tissue cholesterol.

An enzymatic assay currently in use in clinical laboratories for quantitating total serum cholesterol has been modified for the determination of microgram quantities of tissue cholesterol. Two types of assays were developed. In the first, more generally useful method, lipids extracted with chloroform/methanol were solubilized by the addition of detergent and assayed for free and total cholesterol using laboratory prepared mixtures of reagents. Although different detergents were effective, it appears that Triton X-100 may be most universally applicable. In the second type of assay, commercially available reagent mixtures were employed for the determination of total cholesterol only. Results of both types of assays compare favorably with determination using the colorimetric assay on 3beta-hydroxysterols recovered from digitonides.


[Platelets fatty acid variation in patients with atherosclerosis (author's transl)].

Plasma linoleic acid levels were found to be low in the atherosclerosis patients investigated. In contrast, platelet arachidonic acid levels were decreased only when atherosclerosis was combined with diabetes or mixed hyperlipidemia. In acute vascular thrombosis, a marked decrease in platelet arachidonic levels occurrrd, irrespective of whether the patient had atherosclerosis or not.


Subcellular progesterone in human myometrium during late pregnancy.

Samples of myometrium were obtained from twenty-two patients undergoing caesarian sections for various obstetric indications. The myometrium was homogenized and divided into four subcellular fractions (crude nuclear, mitochondrial, microsomal and cytosol). The concentration of endogenous progesterone in these fractions was determined by radioimmunoassay. In addition the concentration of oestrone and oestradiol of fifteen cytosolic fractions was determined. The subcellular concentration of progesterone (pg/mg protein) in human myometrium was higher in samples taken during labour than in samples taken at elective caesarian sections. This finding was statistically significant in the crude nuclear (P less than 0.005) and the microsomal (P less than 0.05) fractions. The absolute concentration of progesterone was lowest in the nuclear and highest in the microsomal fraction. The relative progesterone concentrations in the four subcellular fractions were the same in both groups. The data show that there is no significant decrease in myometrial progesterone associated with labour in man. The mean concentration of oestrone was higher than the mean oestradiol concentration in the cytosol fraction of human myometrium in late pregnancy. This study shows that a completely different oestrogen ratio exists in myometrium than in plasma.


The toxic effect of small iodine supplements in patients with autonomous thyroid nodules.

In sixteen cases of toxic adenoma of the thyroid (autonomous hot nodule with complete suppression of the surrounding normal parenchyma) potassium iodide was given in doses of 100 microgram/day for one week, 200 microgram/day for another and 400 microgram/day for a third week. There was a progressive increase in the serum T4 level. Serum T3 also increased, although this was significant only after the first week. Serum TSH was undetectable throughout the entire period of the study. This metabolic pattern is different from the response seen in cases of nontoxic endemic goitre, where small iodine supplements induce an increase in serum T4 but a decrease in serum T3. Furthermore, the present results may explain the phenomenon of iodine-induced or iodine-precipitated hyperthyroidism (Jod-Basedow) when patients with autonomous thyroid are presented with a high iodine intake. In contrast to the results obtained with small iodide doses, two other cases treated with large pharmacological doses of iodide showed a decrease in both serum T4 and serum T3. It is concluded that the physician should be aware of the possibility of precipitating or aggravating thyrotoxicosis in patients with an autonomous hot nodule by increasing their intake of iodine.


Male pseudohermaphroditism due to partial end-organ insensitivity to androgens.

A 46XY individual with incomplete male pseudohermaphroditism was studied. The phenotype was distinctive, with a masculine habitus despite a small phallus, gynaecomastia and diminished virilization. Plasma levels of testosterone, oestrogens, gonadotrophins and sex-hormone binding globulin were elevated. It is proposed that this syndrome is a variant of Type I incomplete male pseudohermaphroditism.


Triglyceride fatty acids confirm a change in dietary fat.

The pattern of triglyceride fatty acids has been shown to alter rapidly when normalipaemic individuals eating a normal 'western' diet rich in saturated fats were changed to a diet low in cholesterol and high in polyunsaturated fats. The proportion of triglyceride linoleate was used to monitor the adherence to dietary advice of eleven primary hypercholesterolaemic patients (Type II) presenting to a lipid clinic. Patients adhering to diet showed a fall in serum cholesterol and a rise in triglyceride linoleate; recalcitrant patients showed little change. The estimation of triglyceride linoleate may be an important adjunct to clinical judgement in assessing dietary resistance before resource to drug therapy in hypercholesterolaemic patients.


Amiloride in the treatment of primary hyperaldosteronism and essential hypertension.

Amiloride (40 mg/day) was given to nineteen patients with primary hyperaldosteronism. There were significant falls in systolic and diastolic blood pressure, in total exchangeable sodium, and in serum sodium sodium and bicarbonate; while total exchangeable potassium, total body potassium, serum potassium, chloride and urea, and plasma renin, angiotensin II and aldosterone all increased significantly. Amiloride was effective in reducing blood pressure in patients with and without adrenocortical adenoma. No carry-over effect was seen on withdrawing amiloride. Similar changes were associated with amiloride treatment in five patients with essential hypertension; hyperkalaemia was not observed. Only negligible side-effects were encountered in the entire series of twenty-four patients.


The thyroidal production of reverse triiodothyronine in autonomous adenoma.

The ratios of reverse triiodothyronine (rT3) to triiodothyronine (T3) concentrations were similar (0.89 +/- 0.09) in twenty-three autonomous thyroid nodules (fourteen decompensated, nine compensated) and paranodular tissues (0.82 +/- 0.08), whereas the serum ratio was significantly lower (0.098 +/- 0.014). This is compatible with the hypothesis that thyroidal rT3 and T3 production from mono- and diiodotyrosine may be a random process in contrast to non-random extrathyroidal rT3 and T3 formation.


Circadian rhythm of plasma corticosteroids in heroin dependent subjects.

The circadian rhythm of corticosteroid secretion was investigated in six heroin dependent subjects by measuring plasma corticosteroids at 09.00 hours and 24.00 hours. All results were within the normal range and it seems unlikely that chronic heroin dependence affects the hypothalamic control of corticosteroid secretion.


The development of a radioimmunoassay for the measurement of human plasma arginine vasopressin.

A radioimmunoassay for AVP capable of measuring human plasma AVP is described. Iodination was performed by the chloramine T method and purified by chromatography on Sephadex G-25. Specific activity of 125I-AVP was 1710 +/- 155 Ci/mmol. Antiserum of high affinity (Keq = 2.7 X 10(11) 1/mol) has been raised in rabbits, which shows slight cross-reactivity with LVP and negligible reactivity with oxytocin. The aqueous assay is capable of detecting 0.4 fmol of AVP/tube and it is highly reproducible. A F lorisil extraction technique is described in detail and gives recovery of 70% of synthetic AVP added to plasma over a wide physiological range. The lowest detectable concentration of plasma AVP is 0.3 pmol/l. The method has been validated by studying changes in plasma AVP concentration following overnight dehydration (plasma AVP =3.46 +/- 1.89 (SD) pmol/l), and water loading (plasma AVP = 1.54 +/- 0.59 pmol/l), P less than 0.005, in normal subjects. A highly significant positive correlation has been found between plasma AVP and plasma osmolality (r =+0.75). Plasma AVP concentration has also been determined in patients with DI and the syndrome of inappropriate ADH secretion. No effect was found on the level of plasma AVP in normally hydrated volunteers undergoing postural change but levels rose following strenuous exercise from basal concentrations of 1.57 +/- 0.59 pmol/l to 4.77 +/- 3.43 pmol/l, P less than 0.01.


Chromosomal anomalies in patients with retinoblastoma.

Karyotypes from 50 persons with retinoblastoma confirmed by histopathological examination were studied by conventional staining and Giemsa-banding techniques. Two chromosomal anomalies were found. An interstitial deletion of the long arm of No. 13 chromosome was identified by Giemsa-banding in the karyotypes from a boy with unilateral retinoblastoma. Another boy with unilateral tumor had a karyotype of 47,XXY. These findings provide additional evidence that a deletion of chromosome No. 13, most likely involving band 13q14, is associated with the development of retinoblastoma. In conjunction with other reports, our findings also suggest that retinoblastoma may be found more frequently in children with chromosomal aneuploidy.


Tricho-dento-osseous syndrome: a scanning electron microscopic analysis.

A large kindred of which multiple members have the Tricho-dento-osseous syndrome is presented. This is an autosomal dominant disorder characterized by defective enamel, taurodontia, unusually curly hair and occasionally mild to moderate skeletal osteosclerosis. Histologic investigation of teeth (by both LM and SEM) demonstrated that there is a uniformly thin enamel covering with randomly distributed depression and pits. The mineral content of this enamel is closer to that of the underlying dentin, which accounts for its lack of radiographic contrast. The dentin was normal. A bizarre finding is that of a periradicular sheath or membrane that enclosed the open apices and extended partway up the root. It was composed of collagen fiber bundles. The anatomical position of this membrane suggested that it may represent the developing peridontal ligament seen in early tooth formation. Recent embryologic evidence provides support for mesenchymal culpability for all reported features of the syndrome.


Dermatoglyphic patterns in trisomy 8 syndrome.

Up to the present, 38 cases of trisomy 8 have been found. As most of the patients showed mosaicism, the clinical picture is variable and it seems possible that some mosaics will not be detected by the usual cytogenetic examination of blood cultures. We therefore examined the dermatoglyphics of our own case and compared the results with the findings in the other cases reported in the literature, in order to establish a typical dermatoglyphic pattern in trisomy 8 which might be useful in diagnosis. All patients exhibited several unusual dermatoglyphics, including: a low TFRC (x=96.06), high palmar (92.9%) and plantar (100%) pattern intensity, a distally placed axial triradius (62.5%), loop with accessory triradius in an interdigital area (91.7%), thenar (68.2%) and hypothenar (50.0%) patterns, simian crease (47.1%), bilateral arches on the great toes (88.9%) and hallucal-whorl (72.2%). A distinctive feature was the presence of zygodactylous triradii z, z' and z'' (100%) on the soles of the feet, and deep skin furrows on the palms and soles (68.2%). This combination of dermatoglyphic features appears to be characteristic for the trisomy 8 syndrome.


Familial occurrence of chromosome variant 17ph+.

A structural chromosome variant 17ph+ was found in a high genetic risk family. The authors consider the possibility of a causal connection between minor structural changes in the karyotype and congenital defects appearing in one member of the family.


Linking of medical records to form pedigrees.

The human pedigree provides a highly organized representation of historical family morbidity. In order to allow for careful analysis of large numbers of pedigrees, we have developed an interactive computer system for their storage, retrieval and statistical review. The system is capable of integration with our computerized medical record through the use of a common classification (ICHPPC) for patient morbidity. This linkage of pedigrees to clinical data may allow a richer utilization of the clinical experience within Family Practice for the support of research into disease patterns within the family.


Partial trisomy 22: a recognizable syndrome.

A patient identified as being a partial trisomy 22 mosaic is presented. The presence of a translocation t(4;22) (pter;q12) is noted in the mother, sister and maternal aunt. Comparison is made with nine other reported cases of partial trisomy 22 confirmed by parental translocation. These suggest a definite syndrome, including mental retardation, congenital heart disease, skeletal anomalies, anti-mongoloid slant of the palpebral fissures, preauricular skin tags and low-set ears.


Distribution patterns of satellite associations in human lymphocytes relative to age and sex.

Using ammoniacal silver for differential staining of satellites, associations from 1,668 human metaphases were studied from lymphocyte cultures of 167 normal individuals and were correlated with age and sex. There is a peak in satellite association frequency up to age 20 years in males and to 25 in females. With older ages there is a decline in association frequencies. No associations with four or more chromosomes were found in newborns. Furthermore, association in newborns, as well as in individuals older than 50 years, are significantly low compared to in-between age groups. Females have a higher frequency of satellite associations than males between the ages of 3 and 50 years. Typically, females have greater multiple associations, while males have more single associations. Randomness or non-randomness of occurrence of associations fluctuates for different age groups within and between the two sexes.


Insertional translocation into the X chromosome of a 46,XY male.

A baby boy with multiple congenital anomalies and low birth weight has an apparent de novo insertional translocation into the long arm of his only X chromosome. The origin of the translocated material is not evident and both cytogenetic and X-linked marker studies do not indicate any loss of genetic material from the X chromosome. His findings are compared with other males who have X-translocations, none of which appear to be insertional translocations.


Linkage analysis with misclassification at one locus.

It is demonstrated that in linkage analyses with a misclassification error p of the phenotypes at one locus, a wrong specification of p leads to a bias in the estimate of the recombination fraction. In particular, if p is not taken into account at all in the analysis, the recombination fraction is overestimated. This is shown for offspring of phase-known double backcross matings. In the light of these results, the possible effects of dichtomization of quantitative phenotypes are discussed and illustrated with an example of the linkage between hypercholesterolemia and C3.


An autosomal dominant midline cleft syndrome resembling familial holoprosencephaly.

We have detected a previously unrecognized autosomal dominant syndrome characterized by: mental retardation, microcephaly; craniofacial anomalies including cleft lip and anterior cleft palate, hypotelorism and antimongoloid slant; skeletal anomalies, notably of the foot and spine; and chronic constipation. Despite similarities to familial holoprosencephaly, this disorder appears to be a distinct entity. Incomplete penetrance and variable expressivity accompany transmission of the abnormal allele through four generations of a large kindred. Three of the four affected males survived past 20 years of age; the fourth is an infant. All three affected females died very early in infancy.


Bloom's syndrome in two Dutch families.

The clinical and cytogenetic data are presented of four children with Bloom's syndrome, who belong to two unrelated Dutch families. The patients showed, in varying degrees, the clinical features most characteristic of the syndrome: stunted growth; telangiectatic facial erythema; sun-sensitivity of the skin; and decreased immuno-competence. In one child the skin lesions were only minor and the diagnosis would probably not have been made if her sib had not been recognized as having Bloom's syndrome. The cytogenetic characteristics of the syndrome were present in all patients. Each showed a high number of chromosomal aberrations and numerous sister-chromatid exchanges per cell.


Autoimmunity in the relatives of patients with immunodeficiency diseases.

Autoimmune disorders are reportedly more frequent than expected in immunodeficient patients and in their relatives. The hypothesis that genetic factors related to immunodeficiency may predispose to the development of autoimmunity was studied in relatives of patients with variable immunodeficiency (VID), ataxia-telangiectasia (A-T), or X-linked infantile agammaglobulinaemia (X-LA). Close relatives of patients with VID or A-T had thyroid and gastric autoantibodies significantly more frequently than did control subjects. No abnormalities were detected in unaffected relatives of X-LA patients. The increased incidence of organ-specific autoantibodies in close relatives of VID patients was confined to those families with more than one member with immunodeficiency. These data suggest that there are at least two forms of VID, one of which is associated with familial autoimmunity. It is postulated that heterozygous carriers of the A-T gene and persons with genes involved in the development of VID may exhibit T-lymphocyte dysfunction which predisposes them to autoimmunity.


Adverse reactions to nitrofurantoin in relation to cellular and humoral immune responses.

Eighteen patients showing hypersensitivity to nitrofurantoin with symptoms like sudden fever, malaise, cough, pleuritis, leucocytosis and sometimes eosinophilia were analysed for immune reactions, i.e. lymphocyte transformation and specific antibodies. In addition, thirty-three individuals treated with the drug but with no clinical signs of the described reactions were included as controls. The lymphocyte transformation test (LTT) was found positive in ten out of the eighteen sensitive patients while one to three out of fourteen tested control patients were positive, depending on test conditions. An incubation period of 4 days in culture medium supplemented with autologous serum gave stimulation to a greater extent than did isologous serum and/or incubation for 6 days. The antibody determinations, with the enzyme-linked immunosorbent assay (ELISA), showed no IgE antibodies to nitrofurantoin in any of the patients. In contrast, most of the hypersensitive and even the control patients had specific IgG antibodies. The mean value of the antibodies of the sensitive group was higher than of the control group, but considerable overlapping between the groups was noted. In adverse reactions to nitrofurantoin a positive LTT and/or a high antibody titre to the drug favours an allergic mechanism and contributes to a more reliable diagnosis.


Comparison of allergic aspermatogenesis with that induced by vasectomy. II. In vitro studies of cell-mediated immunity to sperm after vasectomy in man and guinea-pig.

Transformation of peripheral blood leucocytes was shown to be a valid assay for cell-mediated immunity to sperm in male guinea-pigs immunized with homologous epididymal sperm (ES) in FCA. A heat-treated extract of ES (BES) was used for culture. Possible developement of cell-mediated immunity to sperm after vasectomy was investigated in patients and guinea-pigs, by culture of blood leucocytes before and at intervals after operation. Patients' leucocytes were cultured with a heat-treated extract of human seminal sperm (BHS); guinea-pigs' leucocytes were cultured with BES. These cultures showed no evidence of specific stimulation of leucocytes by sperm extract, up to 1 year after vasectomy in patients, or up to 6 months in guinea-pigs. Similarly, no evidence of delayed hypersensitivity could be demonstrated by skin tests with BES in animals vasectomized 1 year previously. We consider that this study establishes that cell-mediated immunity to sperm does not develop in either man or guinea-pigs, up to 1 year after conventional vasectomy.


DNA-synthesizing cells in the blood in coeliac disease and inflammatory bowel disease.

The level of 3H-labelled thymidine ([3H]TdR) incorporation of blood cells of patients with coeliac disease was shown in two separate studies to be significantly lower than that of a normal control group. In the first study the 'background' DNA synthesis in unstimulated cultures containing a standard number of blood lymphocytes was measured on days 4, 5 and 6. In the second study a standard volume of freshly drawn whole blood was added to culture medium and the [3H]TdR incorporation measured over the first 24 hr and from 48 to 72 hr. In all cases the [3H]TdR incorporation of cells of coeliac patients on a normal or a gluten-free diet was lower than that of the control group. It is suggested that sequestration of DNA-synthesizing cells in the mucosa of the small bowel may partly explain these results. In whole-blood cultures from patients with inflammatory bowel disease in remission [3H]TdR incorporation over the first 24 hr was raised in Crohn's disease but normal in ulcerative colitis.


C3 metabolism in ulcerative colitis and Crohn's disease.

The metabolism of the third component of complement (C3) has been investigated in four patients with ulcerative colitis, three patients with Crohn's disease and seven control subjects, using radioiodinated C3 prepared from fresh human plasma. Both the fractional catabolic rate and synthesis rate of C3 were increased in the patients with inflammatory bowel disease, although the serum-C3 levels were normal or raised. The results suggest that complement activation may play a role in the pathogenesis of mucosal inflammation in these diseases.


Acute hepatitis: significance of changes in complement components.

Serial measurements of complement components were performed in fifty-nine patients with acute, uncomplicated hepatitis and twelve with alcoholic cirrhosis. Thirty-one of the former group had detectable hepatitis B antigen. Abnormal complement profiles were observed in nine patients with hepatitis B and seven with antigen-negative hepatitis. Low levels of C4, C3 and factor B were common in the subjects with cirrhosis and confined to those cases with severe reduction in serum albumin and/or prothrombin index. By contrast, the complement changes in the patients with hepatitis occurred without significant alteration in these parameters; certain subjects also had reduction in C1q and C5 and a significant number had C3d detectable in fresh plasma. The pattern of abnormality suggests predominant involvement of the classical pathway and it is concluded that this results, at least in part, from an immune process evident only in the early clinical phase of hepatitis. Such gross changes in complement are likely to reflect immune-complex activity and it is proposed that these complexes may be important in the clearance of virus material. The data supports a previous suggestion that recovery from acute hepatitis is primarily dependent on host immune competence rather than viral cytotoxicity or generation of immune complexes.


Asymmetric depression in the serum level of IgG subclasses in patients with nephrotic syndrome.

Thirty-three nephrotic patients were studied for IgG-subclass levels in matched pairs of serum and urinary specimens. Levels in the serum were compared with those found in the sera of non-nephrotic control children and normal adults. Results have shown an asymmetric depression in the serum level of certain IgG subclasses in some patients with minimal-change nephrotic syndrome, focal glomerulosclerosis, and proliferative glomerulonephritis, indicating that the urinary loss of IgG alone cannot account for the low blood levels of this immunoglobulin class in these conditions.


Cellular immune responses to amoebic liver abcess in the guinea-pig.

Guinea-pigs infected in the liver with the Biswas strain of Entamoeba histolytica showed no dermal hypersensitivity but showed positive lymphocyte transformation and macrophage-migration inhibition. The time sequence showed an activated response at 4 days after infection, a full response at 8 days when the liver abscesses were resolving and a waning response at 12 days when the abscesses had healed.


Immune adherence reactivity of rat alveolar macrophages following inhalation of crocidolite asbestos.

The immune adherence phenomenon was used to demonstrate the in vivo deposition of complement on membranes of alveolar macrophages from rats chronically exposed to crocidolite asbestos dust. Pre-treatment of macrophage cualtures with anti-C3 antiserum greatly diminished the level of immune adherence reactivity. Alveolar macrophages exposed to crocidolite asbestos in vitro did not exhibit significant levels of immune adherence reactivity. These results may reflect an in-vivo antigen-antibody-complement interaction on the surface of a alveolar macrophages from animals which have inhaled asbestos dust.


Reduced mitogenic responses in cultures of lymphocytes from newly calved cows.

The proliferative responses to stimulation with phytohaemagglutinin (PHA) in cultures of lymphocytes from newly calved and non-pregnant cattle were compared. Lymphocyte cultures from newly calved cows were seen to respond less to PHA than cultures from the non-pregnant cows. It is suggested that this depression in responsiveness of lymphocytes in the immediate post-parturient period may be involved in the etiology of disease in cattle during the immediate post-parturient period.


A radioimmunoassay for the quantitative evaluation of anti-human acetylcholine receptor antibodies in myasthenia gravis.

A radioimmunoassay was developed for the quantitative evaluation of antibodies to the acetylcholine receptor in the serum of myasthenic patients. AcChR was extracted from human muscle. A detailed preparation of the 125I-labelled alpha-Bgt-AcChR complex used as antigen is reported. Usually, an average of 20 pmol were obtained form 100 g muscle. This preparation is stable for 1 month in presence of an inhibitor of proteolysis and sufficient for performing about fifteen assays. The labelled complex was incubated with increasing amounts of sera and precipitated with anti-human IgG serum. Titres were expressed in pmol 125I-labelled alpha-Bgt-AcChR complex precipitated per ml serum. Out of thirty-nine sera tested thirty-six had positive titres ranging from 0-1-46 pmol/ml. No anti-AcChR were detected in the sera from twenty-seven patients used as controls.


Enumeration of lymphocyte populations in whole peripheral blood with alkaline phosphatase-labelled reagents. A method for routine clinical use.

An improved method is described for the enumeration of lymphocyte surface markers in whole peripheral blood using reagents labelled with alkaline phosphatase. A suspension of washed whole blood is exposed to the labelled reagents and then smeared on slides. Endogenous peroxidase in monocytes is detected by the diaminobenzidine reaction amplified by osmication, and this identifies more cells than are recognised as monocytes by morphological criteria in Romanovsky-stained films. Lymphocytes are identified as peroxidase-negative mononuclear cells and those binding alkaline phosphatase-labelled reagents are demonstrated by treating the smears with naphthol ASMX phosphoric acid and fast red TR salt. By avoiding the loss of lymphocytes which is inevitable in any procedure for isolation of mononuclear cells from the blood, and by permitting elimination of monocytes from the counts, this method enables the proportion and absolute number of different circulating lymphocyte populations to be accurately enumerated. In the peripheral blood of seventeen normal individuals alkaline phosphatase rabbit F(ab)'2 anti-human immunoglobulin stained the following numbers (mean +/- s.d.) of lymphocytes, 9-0 +/- 1-5%, 214 +/- 66/microliter (B cells), and specific rabbit anti-T-cell serum followed by alkaline phosphatase goat F(ab)'2 anti-rabbit immunoglobulin stained 77 +/- 3%, 1846 +/- 488/microliter (T cells). The method, which is applicable to any surface marker which can be detected on living cells in suspension with a soluble reagent, provides permanent preparations which are counted in an ordinary light microscope and permits the use of counterstaining to reveal cellular morphology. Provided that appropriate specific reagents are available it is therefore suitable for routine clinical application.


'Long-distance' lymphocyte study. The effects of transporting blood in lymphocyte blastogenic responses.

A comparison of blastogenic responsiveness to antigens and mitogen by human lymphocytes was made between cells which had been processed for culture immediately following blood collection and cells obtained from blood collected 9-11 hr previously and transported via commercial airline from the patients' homes to our laboratory. There were no significant differences in the responses of transported and non-transported cells if the blood was maintained at ambient temperature during the period of shipment. Chilling the blood during transport, however, resulted both in decreased stimulation of the cells and increased 'background' activity in unstimulated cultures. These findings indicate the feasibility of carrying out both limited immunological evaluations and extended periods of follow-up for patients located at considerable distances from a research laboratory.


Effect of A. laidlawii on murine and human lymphocyte cultures.

Heat-inactivated A. laidlawii (AL) was found to be a potent mitogen for mouse spleen cells. Spleen cells from homozygous nude mice and spleen cells treated with anti-theta serum and complement responded as well as their respective controls, indicating that AL represented a B-cell mitogen for mouse spleen cells. Spleen cells from LPS-unresponsive C3H/Hej mice responded well to AL. The peripheral blood leucocytes from unselected human donors were also stimulated by AL, which appeared to represent a T-cell mitogen for human leucocytes. However, the possibility that it acted as a specific antigen could not be excluded. Attention was drawn to the possibility that the presence of mycoplasma might considerably affect the results of tests where tissue-culture cells or derivatives thereof are added to leucocyte cultures.


Circulating immune complexes in retinal vasculitis.

Seventeen patients with retinal vasculitis, eleven with the peripheral type (Eales' disease) and six with the central type, were investigated to detect the presence of circulating immune complexes (IC) which might then be related to the pathogenesis of their disease. A systemic disease process was identified in six. IC in serum were inferred by the presence of complement (C) activation, rheumatoid factor, Clq or monoclonal rheumatoid factor precipitins, anticomplementary activity, elevated cryoglobulins, inhibition of erythrocyte-antibody (IgG-EA) rosette formation, increased numbers of peripheral blood lymphocytes bearing surface Ig, and spontaneous neutrophil chemotatic activity in plasma. Two or more parameters were positive in thirteen of seventeen patients, with chemotactic activity (69%) and inhibition of EA-rosette formation (59%) being the most frequently positive tests. No immunological differences were detected between the peripheral and central retinal-vasculitis groups. Several IC systems may operate in a give patient.


Ferritin on the surface of lymphocytes in Hodgkin's disease patients. A possible blocking substance removed by levamisole.

Enzymatic radioiodination of surface proteins of Hodgkin's disease peripheral blood mononuclear cells revealed the presence of a blocking protein on their surface. This protein shed into the medium after incubation with levamisole, which resulted in the unmasking of surface proteins similar to those on normal monunuclear cells. The blocking substance was identified. It reacted with anti-human spleen ferritin. It had no detectable iron and dissociated into monomeric subunits of 18,000 mol. wt. by reduction and alkylation, and therefore it is most probably apoferritin rather than ferritin.


Eosinophilia. V. Delayed hypersensitivity, blood and bone marrow eosinophilia, induced in normal guinea-pigs by adoptive transfer of lymphocytes from syngeneic donors.

Dinitrochlorobenzene (DNCB) induced delayed hypersensitivity but no eosinophilia in guinea-pigs from two colonies. Citraconic anhydride (CA) induced delayed hypersensitivity and eosinophilia of the blood and bone marrow, and sites of skin tests were also infiltrated by eosinophils. In adoptive transfer of lymphocytes separated from peritoneal exudate cells of strain XIII-sensitized donors, lymphocytes from DNCB-sensitized guinea-pigs transferred antigen-specific delayed hypersensitivity; lymphocytes from CA-sensitized guinea-pigs transferred delayed hypersensitivity, and induced eosinophilia of the blood and bone marrow of the recipients. Treatment of the lymphocytes before transfer with antilymphocyte (thymocyte) globulin or puromycin suppressed the manifestations in the recipients; normal globulin did not. Active sensitization with DNCB induced formation of small amounts, and with CA larger amounts of anaphylactic antibody. Sera from the actively sensitized animals elicited no significant eosinophilia of blood or bone marrow in one group of recipients. Passive anaphylaxis elicited a transient eosinophilia of the blood, but not of the bone marrow. It is postulated that T-helper cells interact with B-lymphocyte precursors, particularly IgE B cells, to stimulate eosinopoeisis. This results in a reserve of mature eosinophils that may be released in any subsequent anaphylactic event.


Mechanism of complement activation by radiographic contrast media.

Activation of the complement system by radiographic contrast media (RCM) was demonstrated by in vitro haemolytic and immunological assays. Such activation was found to be a function of the RCM molar concentration. Iodipamide was the most active of five RCM tested. When RCM was incubated with normal human serum (NHS) in the presence of ethylene glycol-tetra-acetic acid and magnesium ions, conditions which block activation of the classical pathway but permit activation of the alternative pathway, haemolytically active C3, properdin and factor B were found to be decreased but haemolytically active C4 was normal. Using counterimmunoelectrophoresis, the activation of complement was further demonstrated by detection of C3 and factor B-split products. Finally, when radiolabelled complement proteins were reacted with RCM in vitro and studied by density-gradient ultracentrifugation, it was demonstrated that a large complex was formed with a sedimentation of 22S, similar in characteristics to the C5b-C9 complex. It was postulated that the mechanisms of in vitro consumption of complement by RCM was mainly through the alternative pathway.


Intestinal blood flow in the dog measured by indicator fractionation without tissue samples.

1. A modification of Sapirstein's indicator fractionation technique for measurement of intestinal blood flow has been studied and evaluated in the dog. 2. The concentration of 42K within the intestinal wall was measured simultaneously by a tissue assay procedure and by use of a miniature Geiger-Müller tube within the intestinal lumen. Both methods gave similar results under a wide variety of conditions. 3. Isotope measurement by the Geiger-Müller tube technique obviates the need for tissue samples to be taken and would permit repeat measurements of intestinal blood flow in the same animal.


Prostaglandins: antinociceptive effect of prostaglandin E1 in the rat.

1. The antinociceptive effect of prostaglandins E1, E2 and F2alpha was studied in albino rats. Though all three prostaglandins produced similar degrees of sedation, only prostaglandin E1 (PGE1) produced a dose-related antinociceptive activity. 2. The antinociceptive activities of equi-analgesic doses of morphine (7.5. mg/kg, i.p.) and PGE1 (2.0 mg/kg, i.p.) were inhibited to almost similar extents after pretreatment with drugs known to reduce central turnover of serotonin receptors, namely reserpine, fenclonine (p-chlorophenylalanine), methysergide and 5,6-dihydroxytryptamine. 3. Prostaglandin F2alpha (2.0 mg/kg, i.p.) significantly inhibited the antinociceptive effects of both morphine and PGE1. 4. The prostaglandin synthesis inhibitors, indomethacin and diclofenac, significantly inhibited morphine analgesia. 5. Probenecid markedly prolonged the duration of antinociceptive effect of morphine and the duration of PGE1-induced potentiation of subanalgesic dose of morphine. 6. The results suggest that, in albino rats, PGE1-induced antinociceptive activity is serotonin mediated and that morphine analgesia is not only mediated through serotonin but also through prostaglandins (PGE1 ?) and 5-hydroxyindole acetic acid, the serotonin metabolite.


Chronopharmacology of strychnine and allylglycine in the mouse.

1. The response of individually caged mice to the lethal actions of allylglycine and strychnine was evaluated in animals previously conditioned on an LD 12:12 (12 h light-12 h darkness) schedule in a controlled environment. 2. These convulsant agents were most toxic at 18.00 hours (during the light phase), and least toxic in the dark phase of the programmed lighting schedule. The relationship is considered of the circadian fluctuations in levels of inhibitory transmitter substances to the time-linked action of convulsant agents.


Increased renal sensitivity to aldosterone in the rat: induction by a high potassium diet.

1. The renal response to aldosterone, measured by urinary sodium and potassium excretion, was determined in adrenalectomized rats that had been previously fed either a high potassium diet or a control diet. High K+ rats showed an enhanced response to aldosterone at all doses tested. 2. The induction of this enhanced response to aldosterone could be suppressed by a high sodium intake. It may also require the presence of the adrenal glands during the induction period, although interpretation of adrenalectomy experiments in the present study was made difficult by the effect of maintenance DOC therapy on the renal response to aldosterone. 3. The enhanced response could not be induced by a low sodium diet alone or by the administration of high doses of exogenous aldosterone. 4. No difference between high K+ and control rats could be detected in rat kidney mineralocorticoid receptors, assessed by both in vivo and in vitro binding of tritiated aldosterone. 5. The method of the induction, and the mechanism of the enhanced response remains to be defined.


Nitrogen balance of uremic patients receiving branched-chain ketoacids and the hydroxy-analogue of methionine as substitutes for the respective amino acids.

The efficacy of the D,L-alpha-hydroxy analogue of methionine and the alpha-keto analogues of valine, leucine and isoleucine as substitutes for the corresponding amino acids was tested by measuring the nitrogen balance of chronic uremic patients receiving glycine as their source of nitrogen while eating a virtually protein-free diet. Calcium salts of the following essential amino acid analogues: 2.01 g D,L-2-hydroxy-4-methylthiobutyrate, 2.70 g 2-oxoisovalerate, 3.80 g of 2-oxoisocaproate, and 2.73 g of 2-oxo-3-methylvalerate, plus the remaining five essential amino acids were given to five patients daily for 5--7 days. Nitrogen balance, corrected for urea accumulation (but not for unmeasured cutaneous losses of nitrogen) averaged +1.24+/- 0.39 g/day. We conclude that this mixture and dosage of amino acid analogues can meet the dietary requirements of chronic uremic patients for methionine, valine, leucine and isoleucine.


A method for localizing atrial type B receptors in the dog.

1. The effect of ectopic stimulation of atria (premature ventricular contraction) on the activity of atrial type B receptors has been studied in dogs. 2. In sixteen open-chest dogs, discharge from right or left atrial type B receptors (identified by their response to pulmonary artery occlusion) was recorded. Direct stimulation of either atrium produced an increase in the activity of the left atrial but a decrease in the right atrial receptors. The earlier during the ventricular systole that the premature ventricular contraction occurred the more marked was the effect. 3. In a separate series of fourteen closed-chest experiments the right atrium was stimulated internally via the external jugular vein. The effects on the activity of the atrial type B receptors were similar to those observed during the open-chest experiments. 4. It is concluded that in the dog the technique of internal stimulation of the right atrium without opening the chest can be used to distinguish between the right and left atrial type B receptors.


The significance of vesicoureteric reflux in non-pyelonephritic patients supported by long term hemodialysis.

Vesicoureteric reflux was found unexpectedly during routine investigations before renal transplantation in 12 patients with chronic glomerulonephritis and in one with hypertensive nephrosclerosis. They had all received long term hemodialysis treatment for nine to 106 months (mean 47 months) at the time of micturating cystourethrography (MCU). Four of the patients had previously had a normal MCU indicating that reflux developed after onset of end stage renal failure. The cause of reflux is obscure. It was not related directly to defunctioning of the urinary tract as several patients had daily urine volumes in excess of 300 ml. Infection, another potential cause, was uncommon in patients with reflux. Histology of the excised ureters showed abnormality in most cases with loss of the normal mucosal folds and submucosal cellular infiltrate and fibrosis. These changes are also unexplained. In this group of patients nephroureterectomy for reflux in anticipation of renal transplantation was associated with considerable morbidity. A minimal estimate of the incidence of reflux in chronic glomerulonephritis was 11%. We suggest that in this group and in patients with renal diseases other than chronic pyelonephritis reflux alone does not constitute sufficient indication for nephroureterectomy before transplantation to warrant the risks of major surgery.


Effects of histamine bolus injections and continuous infusions on the H1- and H2-receptors in the hindlimb vessels of the rabbit.

1. Hindlimb vascular resistance (HVR) was continuously measured after pharmacological block of the autonomic effectors in unanesthetized rabbits with previously implanted Doppler ultrasonic flowmeters. 2. Histamine bolus injections caused a dose-related short lived fall in HVR followed by a more sustained rise. The fall was due to H2-receptor stimulation (blocked by burimamide or metiamide) and the rise to H1-receptor stimulation (blocked by mepyramine). At the doses of histamine tested the magnitude of the H1-mediated vasoconstriction had a larger peak effect than the H2-mediated vasodilatation. 3. Histamine infusions up to 200 microgram kg-1 min-1 did not alter HVR significantly but both increases and decreases in HVR were observed after giving H2- or H1-antagonists, respectively. 4. From the double reciprocal plots of 1/peak HVR change and 1/dose of histamine the magnitude of the predicted H1- and H2-mediated peak HVR effects at large doses were the same. This suggested that the number of H1- and H2-receptors were similar in the hindlimb vascular bed, in agreement with the infusion data.


Unusual presentation of tuberculosis in chronic hemodialysis patients.

Four patients developed miliary tuberculosis while undergoing chronic hemodialysis. Two patients had diabetes mellitus. Three of the four patients were hemodialyzed 18--24 months prior to the onset of symptoms. Signs and symptoms included prolonged fever, pleural effusion, pericarditis with pericardial effusion, abdominal pain, weight loss, and ascites. All patients were PPD negative and without historical or radiographic evidence of latent tuberculosis. Disseminated tuberculosis was proven at autopsy in three patients. M. tuberculosis was eventually recovered from pleural fluid and urine in the fourth patient. The immune deficiencies of chronic renal failure and diabetes mellitus are suspected predisposing factors to the development of miliary tuberculosis in these patients.


Studies on normal human skeletal muscle in relation to the pathopharmacology of malignant hyperpyrexia.

1. The effects of dantrolene on pharmacologically-induced contractures and potentiated isometric twitches in normal human skeletal muscle have been studied in vitro. 2. Dantrolene sodium, at concentrations of 3 mumol/l or less, attenuates basal twitch, inhibits halothane potentiation of basal twitch and inhibits halothane-potentiated potassium contractures, but has less effect on twitch potentiation by 2 mmol/l caffeine. 3. Caffeine contractures are attenuated by dantrolene concentrations of 12 mumol/l or greater. The effect of dantrolene on caffeine contracture is characterized by decreased contracture tension and by prolonged time to peak contracture. 4. The results indicate that halothane and 2 mmol/l caffeine have agonistic effects on the excitation-contraction (E-C) coupling mechanism, and suggest that they may act at separate E-C coupling sites. The relationships of these findings to the pathopharmacology of malignant hyperpyrexia are discussed.


Late recovery of renal function in a woman with the hemolytic uremic syndrome.

A case is reported of the hemolytic uremic syndrome (HUS) in a woman taking oral contraceptives. She was treated with heparin, dipyridamole and hemodialysis; and after more than three months, her urinary output rose above 500 ml; and six months after the onset of anuria, dialysis treatment was stopped. This case emphasizes the possibility that HUS in adults is not invariably irreversible and that, despite prolonged oliguria, recovery of renal function can be obtained. Therefore, in adult patients affected by HUS, dialysis should not be discontinued prematurely; moreover, bilateral nephrectomy, for treatment of severe hypertension and microangiopathic hemolytic anemia, should be performed with caution.


Serum dopamine-beta-hydroxylase activity in fasting man.

1. Dopamine-beta-hydroxylase (D beta H) activity was measured in sera from ten healthy male students after 2 and 3 days fasting which is associated with increased circulating noradrenaline. 2. Circulating D beta H remained essentially unchanged throughout the fasting period, although 'stress parameters' like plasma glycerol and free fatty acid concentrations were elevated by a factor of about two from first to second and third day of fasting (P less than 0-01). 3. Serum D beta H activity is not a sensitive index of increased sympathoadrenal activity in fasting man.


Gonadal dysgenesis.

The spectrum of patients with gonadal dysgenesis has expanded over the last decade to include cytogenetically normal individuals. Comprehension of the etiology of gonadal maldevelopment in these patients remains tenuous. More careful study of pedigrees involving 46, XX and 46, XY gonadal dysgenesis may provide better understanding of the mechanism of ovarian failure in these individuals. An important approach will be to identify other disorders occurring in conjunction with primary ovarian failure that can serve as genetic markers for linkage studies. The next decade must provide information that transcends gross structural alterations in sex chromosomes.


Premenarchal vulvovaginitis.

The general subject of premenarchal vulvovaginitis has been reviewed. Vulvovaginal inflammations and infections in the premenarchal child are caused by a large number of etiologic agents. The symptoms, diagnosis, and treatment of the most common of these, namely, nonspecific infections, specific nongonorrheal infections, gonorrhea, protozoal infestations, helminthiasis, mycotic infections, and inflammations due to physical, chemical, and allergenic agents, have been discussed. Reference has been made not only to the older literature but also to some of the pertinent reports published during the last 10 years.


Puberty and menarche.

The various factors that influence puberty and menarche reflect the total environment in which the youngster develops. These influences actually begin in intrauterine life and include genetics, hormones, and nutrition. During the prepubertal years the multifactorial influences on puberty include health, disease, family life, neuroendocrine integrity of the individual, and nltrition. In the female the culmination of the pubertal events is the occurrence of menarche.


Contraception in teenagers.

This review of the special problems of contraception in teenagers has focused on the need for increased efforts in sex education and family-planning and abortion services for this age group. A clear-cut need for new imperatives in these areas is apparent from the data showing an alarming increase in the incidence and rates of unwanted teenage pregnancy and the increased incidence of teenage sexual activity, as compared with earlier times, and its occurrence at earlier ages than previously. Special counseling, education, physical examination procedures, and care and prescription are required when the physician is considering various methods of contraception for adolescents. In the development of adolescent education for responsible sexual behavior, it behooves physicians to keep in mind that it is the members of this age group, in part on the basis of their reactions to the quality of medical care they receive, who will decide and instill the attitudes toward these important issues in the next generation of Americans.


Systemic lupus erythematosus in children: the complex problems of diagnosis and treatment encountered in 101 such patients at the Mayo Clinic.

Between 1945 and 1970, 101 children (86 girls and 15 boys) with systemic lupus erythematosus were evaluated at the Mayo Clinic. Only 9 children were less than 9 years old at the time of diagnosis. The most frequent presenting complaint was arthralgia; fever, fatigue, and a "butterfly" malar rash also were common. Renal involvement, found in more than 76 per cent of patients, was a prognostically poor sign. The overall survival of children with renal involvement is improved by the use of adequate steroid therapy.


Diarrhea, red diapers, and child abuse: clinical alertness needed for recognition; clinical skill needed for success in management.

Three children were chronically given phenolphthalein in the form of laxatives by emotionally disturbed mothers. All children had repeated hospitalizations and extensive diagnostic testing because of recurrent intractable unexplained diarrhea. One child died. Pink-red urine and stool were passed at some time during their illnesses. The presence of phenolphthalein was tested for by application of dilute acid of dilute alkali to the diapers causing respectively disappearance of deepening of the pink-red color. Cases went undiagnosed for prolonged intervals because mothers seemed exemplary. Evidence of their psychiatric illnesses was found once the possibility was considered. Phenolphthalein poisoning should be tested for in any case of intractable diarrhea when objective evidence of intestinal mucosal damage cannot be found. The condition represents a heretofore unrecognized variant of child abuse of battering.


The providing of well-baby care within parent-infant groups. "Pediatricians are encouraged to explore the parent-infant group model in their practices".

A model has been described for the practice of pediatrics using parent-infant groups. In a one year experience, three parent-infant groups met at monthly intervals. A one-hour parent discussion was followed by examination of the infants within the group setting. Among the benefits observed were much more attention of well-baby care and infant development, and the surfacing and alleviation of parental concerns and anxieties.


Effects of delta-9-tetrahydrocannabinol on drug distribution and metabolism. Antipyrine, pentobarbital, and ethanol.

Delta-9-tetrahydrocannabinol (THC) has been reported to inhibit drug metabolism in animals. Twenty-two hospitalized healthy volunteer subjects received THC, 60 to 180 mg/day in divided doses for 14 days. Body weight increased and plasma proteins decreased in all subjects, which is consistent with previously reported plasma volume expansion. Total bilirubin was significantly lower, while other liver function tests remained normal. A within-subject comparison of the pharmacokinetics of antipyrine, pentobarbital, or ethanol given before, during, and after THC was performed. Antipyrine plasma half-life increased during THC in 5 of 6 subjects--mean, 7.9 hr +/- 3.3 (SD) to 9.6 +/- 3.8. Pentobarbital half-life increased in 7 of 8 subjects--mean, 16.9 hr +/- 2.0 to 20.8 +/- 4.2. Blood ethanol disappearance rate decreased in 7 of 8 subjects from a mean of 0.26 mg/100 ml/min +/- 0.05 to 0.23 +/- 0.07. The effect of THC on disappearance rate of these drugs appeared to be due to a combination of: (1) increased distribution volume, due in part to expansion of extracellular fluid volume noted during THC ingestion, and (2) diminished metabolic clearance. THC also delayed absorption of pentobarbital and ethanol in several subjects. This is consistent with THC effects of slowing intestinal motility in animals. The effects of THC on absorption and drug elimination must be considered in evaluating interactions with other drugs.


Effects of actodigin on the heart.

Actodigin is a new semisynthetic cardiac glycoside reported to have a rapid onset and brief duration of action in dogs. Five patients with congestive heart failure in normal sinus rhythm were given incremental doses of actodigin. Overall, there was no significant change in heart rate, aortic or pulmonary artery pressure, systemic vascular resistance, cardiac index, and stroke volume. This lack of response to actodigin is consistent with previous reports of acute administration of other cardiac glycosides. Four patients with atrial fibrillation and a rapid ventricular rate were given similar doses of actodigin. The ventricular rate was readily controlled. After drug administration was stopped, the ventricular rate quickly returned toward predrug levels. Thus, the rapid onset and brief duration of action of actodigin may be useful in the initial management of atrial fibrillation.


Comparative pharmacodynamics of betamethyl digoxin and digoxin in man.

The positive inotropic effects and serum glycoside levels following intravenous and oral administration of betamethyl digoxin (BMD) were measured and compared with those of digoxin in normal adults. The cardiac inotropy was assessed noninvasively in the abbreviation of the electromechanical systolic index (QS2I) obtained by serial measurements of systolic time intervals. Following 0.8 mg BMD intravenously, the QS2I abbreviation occurred within 3 min and reached a maximum at 4 hr. The time-course and the magnitude of the response were essentially the same as those obtained with 0.8 mg digoxin given intravenously in the same subjects. The serum glycoside levels were also similar for both drugs. When administered orally the QS2I abbreviation to 0.8 mg BMD appeared at 40 min and reached a maximum at 4 hr. The time-course and the magnitude of the effects were similar to those observed following oral digoxin. The blood digitalis levels following the drug ingestion, however, were significantly higher for BMD than for digoxin which suggests greater gastrointestinal absorption of BMD. Based on our observations, the inotropic effects of BMD are similar to those of digoxin. Our results, however, do not demonstrate greater cardiac activity after oral administration of BMD despite higher blood levels.


Effect of pentobarbital on the disposition of alprenolol.

Alprenolol was administered orally and intravenously to 5 healthy subjects before and after 10 to 14 daily doses of 0.1 gm pentobarbital. The area under the plasma concentration time curve after an oral 200-mg dose decreased from 706 +/- 277 to 154 +/- 48 ng/ml-hr (mean and SD) with the barbiturate treatment, but there was no significant change in elimination rate. The change in area corresponded to an increase in extraction by the liver from 0.72 +/- 0.13 to 0.93 +/- 0.01. The disposition of a 5.0-mg intravenous dose of alprenolol did not change significantly after pentobarbital treatment. There was no indication of a marked change in hepatic blood flow estimated from the clearance of alprenolol after intravenous administration. It is concluded that pentobarbital administration induces the metabolism of alprenolol in man and that the pharmacokinetic theories derived for hepatic extraction of drugs subject to a high metabolic clearance can be successfully applied.


Drug interactions and multiple drug administration.

Effects of multiple drug administration on adverse drug reactions were studied in 10,518 patients hospitalized on a general medical service during a five-year period. Nine index drug groups, including analgesic, antacid, antiarrhythmic, antimicrobic, anticoagulant, antihypertensive, anti-inflammatory, diuretic, and sedative-tranquilizer drugs, were selected for study. The average number of adverse drug reactions for the anticoagulant and antihypertensive drug groups was higher (p less than 0.05) than for all other drug groups when classified by the number of drugs being taken concurrently (i.e., 0 to 5, 6 to 10, etc.). The rate of reaction for anticoagulant and antihypertensive drug groups was higher (p less than 0.001) than the rate for other drug groups studied. These data suggest a higher risk of adverse drug reactions for patients receiving multiple drugs. The increased risk may result from drug interactions.


Effect of phenobarbital on the disposition of intravenous theophylline.

The disposition of intravenous doses of theophylline was determined in normal male subjects before and after treatment with phenobarbital 2 wk. Although there was some variation in disposition of the 2 drugs, there were no significant effects of phenobarbital on theophylline kinetics. We conclude that theophylline dosage need not be altered during concomitant administration of phenobarbital.


Effects of hexachlorobenzene feeding and iron overload on enzymes of haem biosynthesis and cytochrome P 450 in rat liver.

1. The effect of hexachlorobenzene feeding on liver delta-aminolaevulinate synthase, uroporphyrinogen decarboxylase and cytochrome P 450 was studied at various time-intervals in siderotic and non-siderotic rats. 2 In the non-siderotic group hexachlorobenzene feeding led to a progress decrease in liver uroporphyrinogen decarboxylase activity, accompanied by a progressive increase in delta-aminolaevulinate synthase activity. Cytochrome P 450 concentrations were above normal throughout but fell toward the end of the experiment. 3. Similar but more marked changes were found in the siderotic animals. The fall in uroporphyrinogen decarboxylase activity occurred earlier and was significantly greater in these animals, whereas the increase in delta-aminolaevulinate synthase activity was consistently larger. Liver cytochrome P 450 concentration also rose but to a lesser extent than that in the non-siderotic rats. 4. Hexachlobenzene-induced porphyria would seem to be attributable to inhibition or inactivation of hepatic uroporphyrinogen decarboxylase. Hepatic siderosis has a synergistic effect with hexachlorobenzene on this enzyme and may exert additional effects by promoting cytochrome P 450 turnover.


Effect of pharmacological inhibitors on urate transport during induced uricosuria.

1. The effect on urate excretion of administration of probenecid and pyrazinamide was observed in normal subjects under control conditions and after induction of uricosuria by exogenous urate loading by ribonucleic acid feeding in seven subjects, by volume loading with hypertonic sodium chloride solution infusion in 11 subjects and by partial inhibition of net urate reabsorption with uricosuric drugs in 20 subjects. 2. After ribonucleic acid feeding, the uricosuric response to probenecid was intact and ribonucleic acid feeding did not produce uricosuria after pyrazinamide. 3. After volume loading, the uricosuric response to probenecid was again intact, but infusion of sodium chloride solution still produced uricosuria after pyrazinamide administration. 4. After uricosuric drugs, the uricosuric response to probenecid was diminished. 5. These responses to probenecid and pyrazinamide may reflect different mechanisms of uricosuria. Response to pharmacological inhibitors of urate reabsorption or urate secretion may be useful for classification of induced and clinical uricosuric states.


The association between fasting hyperbilirubinaemia and serum non-esterified fatty acids in man.

1. The concentrations of plasma total and unconjugated bilirubin and of serum nonesterified fatty acids (NEFA) have been measured in two healthy subjects during fasts of up to 21 h. 2. Fasting was either continuous or interrupted by various procedures that altered the concentrations of NEFA and total bilirubin. 3. When NEFA concentrations were increased by the administration of noradrenaline, heparin or caffeine, bilirubin concentrations also rose. 4. When NEFA concentrations were lowered by insulin, bilirubin concentrations fell. 5. Meals of 3-138 kJ and more, taken during the fasting period, lowered total bilirubin and NEFA concentrations in both subjects, whereas the effects of smaller meals were less consistent. 6. These studies demonstrate a statistically significant correlation between total bilirubin and NEFA during uninterrupted fasting and an association between these variables under other experimental conditions. They suggest that the control of bilirubin concentrations in the blood is linked to lipid metabolism.


The variable-pressure neck-chamber method for studying the carotid baroreflex in man.

1. The variable-pressure neck-chamber method was analysed in ten healthy volunteer subjects to determine its suitability for the study of the carotid baroreceptor reflex in man. 2. Positive and negative pressures applied to the neck (range +/- 60 mmHg) were always transmitted linearly to a tissue catheter outside the carotid sinus, but only 86% of positive pressure, and 64% of negative pressure. Tissue pressures were confirmed by simultaneous measurement in the internal jugular vein adjacent to the carotid sinus. 3. Positive and negative pressure changes within the above range did not alter PO2 of internal jugular venous blood, suggesting that cerebral blood flow was unaltered. 4. Positive pressure changes induced reflex pressor responses of similar magnitude at arterial PO2 12-8 and 70-1 kPa (96 and 527 mmHg), suggesting that the carotid chemoreceptors were not involved. 5. It is concluded that the variable-pressure neck chamber is a valid method for selectively studying the carotid baroreceptor reflex in man. However, transmission of external pneumatic pressure to the carotid sinus is imperfect and greater for positive than for negative pressure. This must be recognized to avoid underestimation of gain and distortion of shape of the reflex.


The effect of disuse muscular atrophy on the forces generated in dynamic exercise.

1. Six patients were studied after prolonged immobilization of an injured leg resulting in muscle atrophy. 2. The forces exerted by the atrophied and normal legs during continuous dynamic exercise (one- and two-leg cycling) were examined by a specially adapted ergometer. 3. In one-leg cycling the peak force exerted on the crank at a given work rate, the net work rate performed on the crank, and the proportion of work rate performed in leg extension and flexion phases of the cycle were the same whether the atrophied or normal limb was used. 4. Despite these similarities there was an unexplained reduction in efficiency when using the atrophied leg to perform one-leg cycling. 5. In two-leg cycling the peak force exerted at a given work rate by the atrophied leg was reduced by about 40% as compared with the normal leg, which reflected a similar reduction in the contribution of that leg to the total net work rate. Possible reasons and implications for this disproportionate sharing of work between the normal and atrophied leg are discussed.


Renal clearance of unbound bilirubin in the rat.

1. Unbound or diffusible bilirubin in serum and urine was measured in rats with bile-duct ligation or after continuous infusion of unconjugated bilirubin. 2. The glomerular filtration rate was estimated by measuring the endogenous creatinine clearance and the unbound bilirubin in serum and urine was determined by a Sephadex gel filtration method. 3. Unbound bilirubin clearance was significantly lower than creatinine clearance. 4. The estimated rate of bilirubin reabsorption (calculated from the difference between the unbound bilirubin filtered load and the bilirubin excreted in the urine) was directly related to the serum unbound bilirubin concentration and the unbound bilirubin filtered load. 5. The results suggest that the main mechanism of urinary excretion of bilirubin in the rat involves glomerular filtration and tubular reabsorption by diffusion.


Camphorated and castor oil confusion and its toxic results.

Three cases of camphorated oil ingestion occurring over a 20-month period are discussed. Two of the three ingestions resulted in hospitalization, and the clinical course of the ingestions are presented. Follow-up investigations were performed to determine the causes of the confusion between castor and camphorated oil. Also presented are descriptions of efforts made to improve labeling and packaging of this potentially lethal household product.


A completely automated fluorometric blood cyanide method: a specific assay incorporating dialysis and distillation.

Discrete blood samples can be assayed at a rate of 20/hr. By means of a double lumen catheter, venous or arterial blood can be monitored continuously.


In vitro production of cyanide in normal human blood and the influence of thiocyanate and storage temperature.

Normal human blood stored at room temperature (about 20 degrees C) may, over a period of weeks, undergo a slow transformation of its cyanide content. In contrast, when normal blood is stored at -20 degrees C there is formation of cyanide, usually occurring most rapidly during the first few days of storage. Peak concentrations are never greater than 20 microgram/100 ml, and although some fluctuation in concentration occurs over several months of storage at deep freeze temperature, levels are always higher than in the freshly drawn blood. The amount of cyanide produced appears to be a function of both the initial thiocyanate concentration and the freezing and/or thawing of blood. Blood stored at refrigerator temperature (about 4 degrees C) has the least fluctuation in cyanide concentration over a storage period up to three months. During this study it was found that there is significant difference in whole blood cyanide concentration between smokers and nonsmokers.


The influence of oral penicillamine and diet on lead poisoning in rats.

In spite of continued lead intake, the oral chelate d-penicillamine was effective in reducing blood lead levels of lead-poisoned weanling rats on three different low-residue diets: a standard rat diet, a low calcium diet, and the trace metal salt modified EPA diet. A significant lessening of weight gain was evident in those on the low calcium diet.


Simultaneous identification and quantitation of diphenhydramine and methaqualone.

Methods for the simulataneous identification and quantitation of diphenhydramine and methaqualone in urine have been investigated. Suitable thin-layer and gas chromatographic techniques have been developed. The extraction techniques were evaluated by extracting human urine after the ingestion of therapeutic dosages of the two drugs. A charcoal cartridge technique proved to be superior to direct solvent extraction. Although enzyme hydrolysis gave slightly better recoveries than the charcoal cartridge technique, it is a time-consuming procedure.


Chelation in metal intoxication. IV. Removal of chromium from organs of experimentally poisoned animals.

In an attempt to find more reliable therapeutic or preventive agents for industrial chromium poisoning, some structurally different metal binding agents were screened for their ability to mobilize chromium from liver, kidney, brain, and testes (in vivo) and their subcellular fractions (in vitro), together with the blood corpuscles of experimentally intoxicated animals. Ethylenedinitrilotetraacetic acid (EDTA) and diethylenetriamine pentaacetic acid (DTPA) were quite successful in vivo, while ascorbic acid and 3,4-dihydroxy-L-phenyl alanine (L-DOPA) were more effective in vitro. No relationship could be observed between the structure or the molecular weight of the various chelating agents examined and their ability to remove chromium from either the tissue or their subcellular fractions.


The identification of ketamine and its metabolites in biologic fluids by gas-chromatography-mass spectrometry.

A simple gas chromatography-mass spectrometric method for the identification of ketamine and its metabolites in urine, blood, serum, and plasma has been developed in order to investigate a coma due to overdose of the drug.


Occupational health properties of organic pigments.

Certain terms and statements made by the author of Ref. [1] are criticized. Published evidence dose not support the term "probable carcinogen". "Possible carcinogen" is preferred. Extrapolation of conclusions of carcinogenicity to pigments not yet tested is unwarranted.


RX for social death: the cancer patient as counselor.

Cancer patients frequently encounter psychological problems distinct from those with other serious illnesses. Cancer's deadly nature and its drastic treatment often alienate patients from relatives and caregivers. The CanCervive program trained cancer patients to counsel other patients. They functioned as role models and provided opportunities for catharsis. Yet their own unresolved conflicts often led them to overidentify with patients. Volunteers lacked insight into their own feelings of stress at seeing dying patients and their own guilt over their relatively advantaged medical status. Despite this, they helped patients cope with their insecurities and dependency needs, and assisted with professional education.


Who should be counselors in methadone maintenance programs: ex-addicts or nonaddicts?

Counselors play a central role in most methadone maintenance programs. Yet the choice of who should be employed as counselors is based on little more than personal whims of decision makers. And in evaluating the effectiveness of this highly controlled rehabilitative therapy, the influence of the counselors as an important factor is often ignored. The total changeover of the counselors from an ex-addict to a nonaddict group in an otherwise stable clinic provided an unusual opportunity to compared the performance of the two groups. Both the objective and subjective data supported the hypothesis that ex-addicts as a group compared unfavorably with a similar group of nonaddicts when employed as counselors in a methadone maintenance clinic.


Creating new consultation programs in community mental health centers: analysis of a case study.

A primary prevention program, initiated in a community mental health center, never became fully operational. Analysis suggests that failure to include recipients in initial planning, an unrealistic time table, insufficient institutional support for innovation, the project leader's organizational marginality, and the institutional constraints created by commitment to direct treatment of troubled individuals were factors that contributed to the project's failure. Several recommendations are presented. The most novel and important one is that systems-oriented, preventive mental health work should be based in a separate, distinct institution.