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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.

A simple paper chromatographic method was found to be effective for the study of phenylketonuria and tyrosyluria. It proved to be much more reliable than conventional tests for the detection of abnormal amounts of ketoacids in urine and was suitable and convenient for following the effects of dietary variations on the excretion of these compounds. Information obtained about ketones other than those of primary importance to the conditions studied included confirmation of the excretion of p-hydroxyphenylpyruvic acid in phenylketonuria. A modification of the method applicable to histidinaemia was devised.

In few conditions is the electrophorogram of serum proteins capable alone of deciphering the diagnosis. In conjunction with the assembled clinical data and relevant applied pathological tests the definition of the disease is often clarified. The patterns as observed by the applied pathologist are serially discussed and the importance of the variations in different fractions presented. In several instances, exemplified by liver disease, the assistance rendered toward prognosis has been considerable. Perhaps the most clearcut diagnosis is possible in the agammaglobulinaemia syndrome which is immediately evident by the technique of paper electrophoresis. Current advances are mainly in the direction of estimating the protein-bound carbohydrate and lipid constituents which are believed to reflect the abnormalities in several infections and metabolic diseases. Utilisation of non-ionic detergents has also allowed separation of subfractions of the globulins into numerous new components, including lipo-proteins, some of which had been previously observed during the application of improved buffer systems designed to restrict interference of heavy metals and to facilitate accentuation of fractions having intermediate mobility.

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.

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.

A PAPER Chromatographie study of urinary aminoacids has been carried out in 69 affected and 565 unaffected relatives from 64 families with various types of muscular dystrophy. Only a small proportion of subjects had either generalized or specific hyperaminoaciduria.

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.

According to the result of totally 10 time mass-screening examinations of pulmonary tuberculosis carried out annualy twice spring and fall during five years, the following conclusions were obtained.1) Both the roentogenographically positive finding rate and prevalence rate of actual case had been decreased over a period of five years.2) Cases of mixed form with exsudative and fibroid lesions, according the Oka roentonographic classification, was most apparently decreased in five years. In the forms of the hardnodular and calcified-scar lesions the rates did not any significant changes, and the form with the deformation by surgical operations increased by the spring 1957 and no change was observed after that. Occurrence of exsudative pleurisy was very seldom.3) In the classification of guide standard, number of the patients with the A calss (off work required) and the class B (limitted working hour) remarkably decreased, and those of the class C (under supervision) relatively increased.4) According to the classification by the Japan Society of Tuberculosis, no case with extensively advanced cavity form was found through the observation, and cases with not-advanced cavity form disappeard by the end of the observation. Also cases with the varied non-cavity form was obviously decreased. No apparent changes were shown in the unmber of cases with the stable non-cavity form and the recovered form.5) The decrease of the number of cases with exsudative form seemed to be more apparent in the group of workers with medium physical work, compared with that group of desk workers. The increase of both the roentogenographically positive finding rate and prevalence of actual case was observed by the older years of age, and the trend was not so much varied through the five year observation.

The action of heparin and heparinoid substance (sulfated chitosan) on the cholesterol-fed rabbit serum was as follows:1. An increase of the Sf 30-70 component and a decrease of the Sf

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.

Whole-cell hydrolysates were prepared from 58 strains of nocardiae and streptomycetes. Strains morphologically intermediate between the two genera and morphological variants of the same strains were included. Paper chromatograms made from the whole-cell hydrolysates clearly demonstrated meso -diaminopimelic acid as a major constituent of cultures of Nocardia spp., and LL-diaminopimelic acid as a major constituent of cultures of Streptomyces spp. In cultures of ten strains of N. madurae and of three of N. pelletieri, meso -diaminopimelic acid predominated, thereby supporting the assignment of these species to the genus Nocardia .

When purified diphtheria toxoid was subjected to filter paper electrophoresis a separation was achieved under certain conditions. The flocculating activity of the toxoid would appear to be localized in only one of the four fractions obtained. This fraction represented about one-third of the material used (0.05 ml.), thus indicating the relative impurity of a highly purified material.

Dahlqvist, A. Test Paper for Galactose in Urine. Scand. J. clin. Lab. Invest. 22, 87-93, 1968. Galactose excretion in the urine is an important symptom of hereditary galactose intolerance (galactosemia). Demonstration of galactose in urine with galactose oxidase would offer a more specific method of diagnosis of this disease than the methods which are currently being used.Urine contains inhibitors, however, which complicate the determination of galactose with galactose oxidase. The properties of these inhibitors have been studied. Their properties are similar to those of reduced glutathione, which substance may play an important role.A galactose oxidase test paper for galactose in urine is described in which the inhibitors are removed by adsorption before the sample reaches the colour reagent part. The test paper has high sensitivity for galactose, but does not react with glucose, lactose, or galactitol.

Since blood spots on filter paper are routinely collected in many areas for phenylketonuria testing of newborn infants, we developed several methods for detecting sickle-cell disease and other hemoglobinopathies using this type of specimen. Simple, inexpensive modifications of cellulose acetate electrophoresis plus citrate-agar and globin-chain electrophoresis are employed, permitting one technician to screen as many as 500 specimens per day at a material cost of $0.03 per specimen. (N Engl J Med 288:1265–1268, 1973)

1.1. The results obtained by paper electrophoretic analysis of the serum protein distribution of 1,516 admissions to an army general hospital are described. The differences between results obtained by paper electrophoresis and by moving boundary electrophoresis, and the reasons for these differences are discussed briefly. 2.2. Infections (particularly tuberculosis), malignant neoplasms, arteriosclerosis, rheumatic heart disease, hepatitis, cirrhosis, rheumatoid arthritis and sarcoidosis were most frequently associated with abnormalities in serum protein distribution. 3.3. The most repeated abnormality in this series of cases was a decrease in the serum albumin concentration, followed by elevation of the alpha 2 globulin, gamma globulin, beta globulin and alpha 1 globulin. Increased levels of albumin and decreased levels of the serum globulins occurred rarely. 4.4. The serum protein distributions of 325 women and of forty-one newborn infants are described. 5.5. The results of paper electrophoretic analysis of the serum proteins may be pathognomonic for multiple myeloma, nephrosis and hypogammaglobulinemia, and useful in the diagnosis of hepatitis, cirrhosis, tuberculosis, sarcoidosis, kala-azar and lymphogranuloma venereum. In addition, information may be obtained regarding the state of disturbed physiology, the activity and, occasionally, the diagnosis in such conditions as rheumatoid arthritis, rheumatic fever, malignancy, sequelae of arteriosclerosis and infectious diseases.

The fluorescent antibody technique first described by CooNs, CREECH and J o ~ s (1961) has been widely used for the detection of antigen and antibody in numerous systems including infectious diseases where it has found its widest and most important use from a public health standpoint. BOZFMAN and ELISBERG (1963 and 1967) applied the technique to the serological diagnosis of scrub typhus and their procedures are now being used in laboratories throughout the world. The technique described by them is one of the most reliable serological tests known; however, it requires collection of a volume of whole blood which is not always possible in epidemiological work or in small mammals. Other workers have encountered similar problems with other diseases and ANDERSON, Sta)trN and WILLIAMS (1961) reported on a method that used dried whole smears on filter paper in the fluorescent antibody test for the diagnosis of schistosomiasis. More recently, BAILEY, CUNNINGHAM and KIM~ER (1967) reported on the same technique adapted for a study of trypanosomiasis in Africa which Walton and Arjona reported on its adaption for use in the indirect fluorescent antibody test for toxoplasmosis. This paper reports on the combination of these two techniques for the detection and titration of antibodies against the causative agent of scrub typhns--R, tsutsugamushi.

A rapid screening method for detecting glucose-6-phosphate dehydrogenase (G6PD) deficiency is described. The method involves the collection of blood samples on filter paper and depends on the fluorescence of reduced nicotinamide adenine dinucleotide phosphate. Simple and inexpensive to perform, this test yields positive results when enzyme levels are less than 25 per cent of normal, thus permitting the easy detection of hemizygous men and homozygous women with G6PD deficiency. Also, this test may give positive results in some heterozygous female carriers.

The paper-strip blood-sampling technique was evaluated for efficacy in plague passive hemagglutination tests. It is valuable for widespread serological surveys.

A fully automated two-channel filter paper (FP) hemagglutinating machine was developed for the detection in prenatal sera of the large majority of antibodies related to hemolytic disease of the newborn. In the tests of 9738 first trimester sera, 50 with anti- Rh , one with anti- rh ′, three with anti- rh ″, and four with anti- hr ′ were found. Parallel studies of the prenatal sera with sensitive manual methods and with an automated colorimeter-recorded (CR) machine gave equivalent results. The productivity of the FP machine compared with manual methods and its simplicity relative to the CR machine indicate that the FP machine is satisfactory for the purpose for which it was developed.

I 'HIS report describes a case of dermatitis due to contact with the 4-tertiary butyl catechol in Thermo-Fax paper as it was manufactured before 1958, as well as the methods employed to identify ...

A glucose-specific test paper which is sufficiently sensitive to react with the small amounts of glucose present in urine from normal persons in the fasting state can be used to discriminate between urine samples containing more than and those containing less than 1 to 1.5 mg of glucose per 100 ml of urine. This provides a screening method for significant bacteriuria in an apparently healthy population since urine samples, obtained under standardized conditions from subjects in the fasting state with such bacteriuria, contain either very little or no glucose.

Proteinuria, edema, and an increase in the diastolic pressure, which occurs in the latter months of pregnancy, are well-established indications of preeclampsia. Of these findings, consistent proteinuria is considered most important because it is usually detectable before other signs of preeclampsia appear and because the quantity of protein excreted generally parallels the severity of the condition. Success in the management of the preeclamptic patient is markedly enhanced by its earliest possible diagnosis. Consequently, it is clinically essential to test for proteinuria frequently. It is our routine practice to see all uncomplicated obstetrical patients in the office or the clinic at monthly intervals during the first two trimesters, every three weeks during the seventh month, every two weeks during the eighth month, and weekly during the final month. Testing for proteinuria during these visits is done routinely. However, patients do develop toxemia, sometimes to a marked degree, between visits. For about

A paper chromatographic analysis in 60 patients with muscular dystrophy from 49 families and in 351 of their relatives showed a high incidence of generalized hyperaminoaciduria in both patients and relatives. There was a higher incidence of threoninuria in patients and their relatives than in controls. One family was found to have excess histidinuria and another excess lysinuria.

Lyphan paper M 25 can be used for pH measurements in milk and whey in the pK range 5·4—6·6. As a rule the error does not exceed 0·1 pH.In very acid milk and whey (pH < 5·4) lyphan paper of the L series may be used. Generally the deviation does not exceed -0·2 pH. In alkaline milk and whey lyphan paper cannot be used.In milk and whey from cows infected with mastitis M 25 may be used unless the pH exceeds 7.

Protein antigens fixed on paper in the form of azocompounds or by simple drying at room temperature have a high serological activity and specifically fix the complement in the presence of a corresponding anti-serum. This was established by the method of quantitative reaction of complement fixation (50% titer). Complement fixation test (50% titer) for “paper” antigens gives an opportunity of using different antigens in the same test and replacing them during the experiment. This permits the use of antigens adsorbed on paper and the quantitative complement fixation test for investigation of the optimal conditions of obtaining the specific antisera.

Amino-acids in urine were analyzed by means of paper partition chromatograph, both clinically and experimentally.Usually two-dimension method is performed, but it is also of some clinical value to detect amino-acids with large Rf by the use of one dimension method.Alanine, Glycine and Taurine always appear in urine, no matter whether it be normal or pathological.As a rule, amino-acids detected in urine of hepatic patients increase in number; especially in the jaundice-stage of acute hepatitis there can be ascertained several sorts of amino-acids, while in the reconvalescence they decrease in number showing tendency to become normal.In cases with liver cirrhosis, detected amino-acids are of various number, only the appearance of amino-acids with large Rf is more remarkable than in those who suffer from hepatitis. This occurrence is neither concerned in existence of icterus nor ascites.In cases with hepatic coma, some increase in number very much, others not so markedly. That is to say, patients suffering from hepatic coma do not always excrete high-degree amino-aciduria.Number of amino-acids proved in hepatic cancer is relatively few, and there is no great difference between in cancer and innormal.Number of amino-acids detected in cholecystitis and cholecystopathy fluctuates instablely. In above mentioned cases there can not be proved any special amino-acid appearing without fail.Attitude of amino-acids in urine of rats which livers were damaged with various kinds of toxins, is similar to that of clinical cases.High protein diet after administration of hepatic toxins is admitted as effective to prevent the appearance of a certain amino-acids in urine.

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.