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A method is described for rapid analysis of phenylalanine and tyrosine using eluates of whole blood that has been impregnated into filter paper. This method employs semi-automated ion-exchange column chromatography (amino acid analyzer) and will yield reasonably accurate and consistent quantitative values for phenylalanine and tyrosine concentrations within two hours. The technique has significant value in rapidly confirming the presence of increased phenylalanine with or without a concomitant increase in tyrosine concentration in autoclaved or unautoclaved blood filter paper specimens. Thus phenylketonuria can be differentiated from hyperphenylalaninemia due to tyrosinemia within hours after the initial identification of an elevated phenylalanine concentration by a screening method such as the Guthrie test. In addition, further evaluations of phenylalanine and tyrosine concentrations can be facilitated in infants on whom only blood filter paper specimens are obtainable.

1.1. Filter paper electrophoretic analysis of the serum and urine proteins in thirty-five cases of multiple myeloma has been carried out. In all thirty-five cases characteristic protein abnormalities were observed; seventeen cases showing homogeneous peaks in both the serum and urine, eleven cases in serum only, seven cases in urine only. 2.2. All urine samples showing a positive Bence-Jones protein reaction also displayed a characteristic electrophoretic abnormality. Eight of the twenty-four electrophoretically positive urine samples were Bence-Jones protein negative by the usual tests. Because of the relative simplicity of urine electrophoresis by the filter paper technic, this test is considered to be a practical clinical procedure. 3.3. Two cases are presented in detail because certain atypical clinicopathologic features of their disease patterns possibly assist in an understanding of the relationship of the plasma cell dyscrasias to chronic lymphatic leukemia and the lymphosarcomas. A third case, which demonstrates the value of serial electrophoretic studies in myeloma, is also presented. 4.4. Consideration is given to some of the physicochemical properties of the myeloma serum and urine proteins, and to the possible nature of their interrelationship.

Filter paper electrophoretic analyses of serum were carried out in a series of eighteen proved cases of multiple myeloma, twenty-three normal controls and 130 patients with non-myelomatous diseases, and compared with the results of classical Tiselius electrophoresis and chemical fractionation. Abnormal electrophoretic patterns were found in all of the cases of multiple myeloma studied, and tall narrow spikes representing abnormal proteins with varying mobilities in the globulin range were demonstrated in 78 per cent. The latter were considered diagnostic of myelomatosis since similar patterns have not been described in any other disease and since none were observed in any of the present controls. In six of the eighteen cases the diagnosis was first established on the basis of these electrophoretic patterns and subsequently confirmed by bone marrow biopsy. In eight others they were merely confirmatory in previously diagnosed cases. Of special interest were the diagnostic patterns found in four patients in whom myelomatosis was not suspected. There were significant quantitative differences between the results obtained by filter paper and classical electrophoresis, and the results of both differed from those of chemical fractionation. However, the diagnostic peaks of myelomatosis were equally well demonstrated by both electrophoretic methods. It is concluded (1) that the electrophoretic patterns of serum obtained by both the filter paper and Tiselius methods exhibit highly characteristic peaks in a high proportion of individuals with multiple myeloma, (2) that such patterns are of great diagnostic value and (3) that the filter paper method offers the advantages of simplicity, low cost and speed, without any loss in sensitivity or specificity, which make it especially valuable for screening large numbers of specimens in patients suspected of having myelomatosis.

By the method of paper electrophoresis a comparative quantitative study was made of the sundanophilic serum lipids of two groups of persons; one group consisted of twenty-one patients who had had previous myocardial infarction, the other of eighteen women with no clinically important arteriosclerosis. The electrolipograms in all cases revealed two zones, the larger probably corresponding to β-lipoproteins, the smaller to α-lipoproteins. In the atherosclerotic group the major lipid band constituted a larger fraction of the total lipid than in the control group. The major lipid band of one-third of the atherosclerotic persons extended farther with respect to the β-globulin.

Separations of microgram quantities of radioactive mercury, thallium, bismuth, and lead, and of non-radioactive platinum, gold, copper, nickel, cobalt, iron, cadmium, silver, and palladium, in from 2 to 20 min, have been achieved using a new method of ascending paper chromatography with acetone containing various complexing agents as the developer. The new technique involves the use of thick absorbent paper having high wettability, a developer of high wetting power, and, in particular, an atmosphere which is supersaturated with vapours of the solvent.

Summary A qualitative method of paper chromatography for phenolic compounds is discussed. Whatman No. 7 paper was impregnated with a saturated solution of boric acid. The solvent used was the organic phase of n -butanol saturated with 2% NH 4 OH. The time of development was 15 hours. Beside the classical methods of spot detection, two new processes were employed: (a) ultra-violet examination of the chromatogram after Maule's test; (b) spraying with a solution of diazotized sulphanilic acid, exposure to ammonia vapours followed by spraying with a solution of diazotized p -nitroaniline. A table of R F values of 34 pure phenolic compounds is given, together with methods for their identification.

Human sera were absorbed into electrophoretic paper (Whatman) discs. After drying and storage either at +4 °C or at room temperature (18 °C) for four, seven and 14 weeks, the sera were eluted with distilled water and the diphtheria antitoxin contents were determined. Comparison with the titration of sera stored in the conventional way showed that there had been a significant decrease in titre in the case of discs dried and stored at room temperature. Alternatively, when stored at +4 °C a significant decrease in titre did not take place. This method of storage can thus be used where deep freezing of serum is not available. The storage and the transportation of sera treated in this way is very convenient.

The electrophoretic patterns of the sera of 25 patients with sarcoidosis were compared with the patterns obtained in 31 persons suffering from other diseases. Only in the cases of sarcoidosis were we able to find a characteristic mean electrophoretic pattern that is a increase in the alpha 2, beta and a marked increase in the gamma globulin while the albumin showed a decrease.

Use of the fluorescent antibody test for the laboratory diagnosis of human schistosomiasis on minute amounts of dried blood specimens is described. Sensitivity and specificity of the test appear to be comparable to the best described serological tests. Mailing of specimens under adverse conditions did not seem to influence the results. Determinations of antibody titers were possible. These findings and the ease with which specimens can be obtained, delivered to the laboratory and tested suggest that the procedure may be useful in epidemiologic surveys offering some advantages over the known methods of stool examinations, serology, and intradermal testing.

The development of rapid and dependable methods of paper electrophoresis for fractionation of serum proteins has brought a valuable contribution (1) to diagnostic laboratory procedures. Electrophoretic analysis in the pemphigus group of diseases has been made in the search of yet They generally agreed that in the acute form and in the extensive and severe cases of the disease the total protein and the albumin were decreased and the gamma globulin was increased. The results for the other fractions showed some discrepancies. The chronic or mild cases were TABLE I Number Age (years) Duration of Disease Last Shower of Bullae Sedimentation Rate (Wester- gren-mm/ hour) General Condition Albumin (%) Globulins Total Protein Albu-of Patients Alpha-l (%) Alpha-2 (%) Beta (%) Gamma (%) G?ob ulin 1 23 4 ys.

A mixture of thiouracil, methyl- and propyl-thiouracils can be separated by paper chromatography with amyl alcohol/water and the spots shown up with I 2 vapours or by spraying with dichloro-benzoquinone chloroimide and alkali.

Summary The study of various forms of mellituria has been neglected by the clinician for technical reasons. One-dimensional paper chromatography is a suitable new technique for extending our knowledge of this subject, as it is both sensitive and specific and can be performed in any clinical laboratory. The method we have used for the last 8 years is described in some detail. The sugar excretion in 170 twenty-four-hoururine collections of 104 full-term and premature infants has been examined and a physiologic mellituria has been observed with a moderate increase of glucose, galactose, fructose, and lactose, which lasts for the first 10 to 14 days. The pattern of the mellituria points to immature functions of the renal tubule, the liver, and the intestinal tract. There is no close time relation to the physiologic aminoaciduria of infancy. In 125 healthy children and adults, urinary excretion of sugar was minimal or absent. To obtain a general idea of the urinarysugar pattern in various diseases 1,000 selected patients were investigated, 130 of whom showed mellituria. In 114 instances the mellituria was due to the increase of a single sugar, namely to glucose in 77 cases, lactose in 13, fructose in 12, galactose in 11, and xyloketose in 1, respectively. In 16 cases the mellituria was due to an increase of more than one sugar. The diseases associated with these melliturias are discussed and some tentative figures are given as to the frequency of this association. Isolated and constant melliturias are characteristic of inborn errors of sugar metabolism while symptomatic melliturias are more variable in pattern and occurrence, though different sugar patterns can be distinguished in hepatic, renal, and intestinal disorders. The sugar excretion in the urine is affected by the sugar intake. To establish standard test conditions a sugar loading test was devised by mixing 40 Gm. glucose, 8 Gm. galactose, 30 Gm. fructose, 8 Gm. lactose, and 20 Gm. sucrose into a sugar “cocktail.” This amount was given to adults and two-thirds or one-half of it to children. Urine was collected in the fasting state and at 1, 2, and 3 hours after the sugar load. Although mild mellituria occurred after the load in every one of 45 healthy controls, the sugar increase was more pronounced in diseases of the liver, kidney, and intestine. The diagnostic possibilities of such a loading test are underlined by the fact that the fasting urine repeatedly failed to show mellituria. There was a poor correlation between the chromatographic results and Benedict's reduction test. Sugar concentrations up to 150 mg. per 100 ml. were associated with blue or blue-green Benedict reactions, and green reactions repeatedly occurred in urines free from sugar. Definite and diagnostically important melliturias such as idiopathic galactosuria may be missed if clinical investigations are limited to reduction tests, even as a screening procedure. For the accurate diagnosis of mellituria, paper chromatography is recommended as a routine procedure.

One drop of a solution of the unknown substance in hydrochloric acid is run by the ascending method on paper with ethanol-hydrochloric acid-water as solvent for about 20 hr at room temperature. Subsequently, the strip is sprayed with a 0.2% solution of morin in ethanol. The chromatogram is viewed after drying in this “acid” state and again after treatment with ammonia vapours, both in reflected and in transmitted sunlight and ultraviolet light. From the appearance, shade of colour, fluorescence and location of the spots visible under different circumstances, conclusions can be drawn as to the composition of the sample.

In recent years several papers have appeared describing the analysis of serfim proteins in bullous diseases. Most of the work has been carried out on serum from casesof pemphigus. Lever et al. (1) showed that in pemphigus vulgaris there was a decrease of serum albumin and an increase in a\ , a2 , /? and 7 globulins. They indicated that the increase in a\ and 2 globulins could be attributed to increase in glycoproteins. There was a tendency after treatment with steroids for the albumin fraction to increase and for the ol\ and a2sb> fractions to return towards normal although still remaining slightly elevated.

Summary Attention has been directed to the need for constant vigilance in safeguarding young infants from accidentally ingesting small foreign objects that may become lodged in the esophagus. Prompt dislodgment and removal of these objects may avoid dire complications. If undiagnosed, their lodgment in the esophagus may give rise to respiratory symptoms and findings of a bronchitis unresponsive to any medication. This pulmonary involvement is dissipated after removal of the foreign body. The essentiality of roentgen-ray examination of the chest even in young infants with unexplained respiratory symptoms cannot be under estimated in the early detection of a foreign body arrested in the esophagus. In our case it was conjectured that the accidental ingestion of the foreign body (paper clip) that became lodged in the esophagus might have occurred during the neonatal period, whence the respiratory symptoms had their onset.

A case of a carrier of two albumins in the urine is described. The percentual values of serum and urinary proteins obtained by paper electrophoresis are presented. The results of this analysis demonstrated a genetic defect accompanied by an evident nephrotic syndrome that, in the final stages of the disease, allowed complete passage of the albumin into the urine. Using an improved resolution method, buffered and un-modified urines were studied. The electropherogram of the ascitic fluid was similar to that of serum. Other complementary tests were also carried out. Comparing the relative percentages of normal and abnormal albumins (discounting in the latter the value of α 1 -globulin) it can be seen that each had a value of approximately half of the complete albumin fraction.

A modified version of the Hochella [2] automated fluorometric technique for tyrosine determination is presented as a mass screening procedure for tyrosinemia. With this technique, 85 000 tests a year can be done with one Auto-Analyzer. In the province of Quebec, 17 cases of hereditary tyrosinemia have been detected with this method over the past three years (208 000 tests). Although the number of false positives is high, blood tyrosine determination remains, to our knowledge, the only mass screening procedure available to detect hereditary tyrosinemia.

SUMMARY The performance of three haematological diagnostic keys (Bendixen, Tolle and Theilen) for the pre-tumorous form of bovine leukosis was studied in Chilean cattle. In all categories, Tolle’s key showed the lowest number of animals classified as positive or suspect for leukosis and the highest number of animals with persistent lymphocytosis. Animals affected with brucellosis were frequently positive for pre-tumorous forms of leukosis.

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Ninety-four aspiration lung biopsies were performed on 73 patients. These were divided into the four groups shown in Table I. Thirteen biopsies were done on normal lungs to show the histology and bacteriology (Group I); there were 29 biopsies of cases where the diagnosis was uncertain (Group II); the pathogenesis of the resolution of acute pneumococcal lobar pneumonia was investigated by means of 2? biopsies (Group III); and the pathogenesis of chronic lobar pneumonia by means of 20 biopsies (Group IV). In addition, hepatic cells were obtained in four biopsies and splenlc cells in one. In all, seven cases were done after death but the term “biopsy” has been retained for the sake of convenience. Tables II and III show the sex, race and age distribution of the 73 patients. Most of the biopsies were done at the New Somerset Hospital (Cape Town) which is exclusively for non-Europeans, and thus the majority of the patients were Colored or African. The oldest living patient to be biopsied was Case 5, a colored woman of 74 years; the youngest was Case 68, a colored male infant of four months. The cases were numbered according to the order in which they were done.

1. The three leading surgical approaches to the revascularization of an ischemic myocardium are those of: a.) Beck: heart surface abrasion; insertion of coarsely ground asbestos; occlusion of the coronary sinus to 3 mm.; and application of the pericardium and mediastinal fat pad to the surface of the heart; b.) Thompson: insertion of magnesium silicate, a non-absorbable and irritating substance into the pericardial sac. c.) Vineberg: implantation of the left internal mammary artery into the left ventricle via a myocardial tunnel. 2. Noteworthy and provocative animal experiments to show the validity of revascularization procedures have been carried out by: a.) Beck, who has shown that his technique adds a flow of 4.7 c.c. of blood per minute to an ischemic area of myocardium, and that the quantity of backflow decreases the mortality in dogs whose coronaries were subsequently ligated; b.) Vineberg, who has demonstrated anatomically functioning vessels of arteriolar size between the coronary circulation and the extracardiac source, these anastomoses protecting the animal from a myocardial infarction post ligation of one of the major coronaries. He has also artificially recreated the gradual occlusive process of atherosclerosis, and by exercise tolerance tests has shown that animals receiving his procedure midway in their ischemic decline from normal tolerance regain most of their exercise potential, whereas the controls continue to lose their original exercise tolerance. 3. The human results of coronary heart disease surgery are promising: there has been a substantial decrease in the pain of angina pectoris, and an increase in the work capacity of the coronary heart disease patient; and there is some evidence that the eventual life expectancy of patients undergoing surgery has been increased. 4. It is evident that the greatest difficulty in justly appraising the benefits of surgery to the coronary heart disease patient is the lack of proper control series anywhere in the literature.