Showing papers in "Journal of Clinical Pathology in 1954"

Estimation of Plasma Phosphatase by Determination of Hydrolysed Phenol with Amino-antipyrine

Abstract: Plasma phosphatase was first determined by Martland (1925) and then by Kay (1930), using glycerophosphate and estimating the inorganic phosphate liberated at 370 C. at a specified pH and in a given time. The procedure has been variously modified and simplified by Jenner and Kay (1932), Bodansky (1933), Shinowara, Jones, and Reinhart (1942). Several substrates, other than glycerophosphate, have also been used, e.g., phenyl phosphate (with determination of either the liberated phenol, King and Armstrong, 1934, or phosphate, King, Abul-Fadl, and Walker, 1951), nitro-phenyl phosphate (by the colour of the liberated nitrophenol, Ohmori, 1937; King and Delory, 1939; Bessey, Lowry, and Brock, 1946), naphthyl phosphate (by determination of naphthol by a diazo method, Seligman, Chauncey, Nachlas, Manheimer, and Ravin, 1951), phenolphthalein phosphate (by the colour of alkaline phenolphthalein, Bray and King, 1943; Huggins and Talalay, 1945). Of the.se procedures, the Bodansky (glycerophosphate) and the King-Armstrong (phenyl phosphate) are perhaps the most widely used. King and Armstrong (1934) determined free phenol, liberated from phenyl phosphate by the phosphatase, by either the Folin and Ciocalteu (1927) phosphomolybdic reagent or by the diazo reaction, and preferred the former. Both procedures required the precipitation and removal of the plasma proteins subsequent to the hydrolysis of the substrate. In 1951 Grifols Lucas proposed a procedure for determination of the free phenol, which did not require removal of the proteins. This was based on the 4-amino-antipyrine reaction of Gottlieb and Marsh (1946). In this method the conditions under which enzymic hydrolysis is carried out are the same as in the King-Armstrong method, as modified by King (1951), the amino-antipyrine reagent (A.A.P.) being used to estimate the phenol liberated. 4-Amino-antipyrine reacts with certain phenolic substances in the presence of alkaline oxidizing agents to produce quinonoid substitution products. These give a red colour proportional to the phenol present, and can be determined colorimetrically. Grifols-Lucas found that A.A.P. does not react with plasma proteins, and their removal is unnecessary when phosphatases are estimated.

Serum 5-Nucleotidase

Abstract: REIS1–4 has demonstrated the presence of a phosphatase in various tissues specifically hydrolysing 5-nucleotides, for example, adenosine-5-phosphate. One of us has previously reported5 that normal blood serum has a low 5-nucleotidase activity corresponding to about 1/10 of its non-specific alkaline phosphatase activity towards glycerophosphate. In bone diseases where serum alkaline phosphatase is raised, serum 5-nucleotidase may be normal; but in jaundice, either infective or obstructive, both enzyme levels may be increased, those of the 5-nucleotidase sometimes exceeding those of the non-specific alkaline phosphatase.

The determination of serum acid and alkaline phosphatase activity with 4 - aminoantipyrine (A.A.P.).

Abstract: C6H5 Gottlieb and Marsh (1946), using alka:ine ferricyanide as the oxidant, devised a method for the estimation of certain phenolic fungicides with A.A.P. The procedure was modified by Grifols (1951) for the determination of phenol in the measurement of alkaline phosphatase activity in body fluids. This author made a careful study of the factors which control the development of the colour and his results showed good correlation with those obtained by the method of King (1951). Advantages claimed for the method were that the reagent did not react with proteins, thus obviating their precipitation during the estimation, and also the colour development was comparatively rapid. The use of A.A.P. as a reagent for phenol appeared to be of potential value especially if its use could be extended to the estimation of serum acid phosphatase activity. Considerable modification of the original Grifols procedure was found to be necessary in order to measure both alkaline and acid phosphatase activity in blood. The use of calibration curves, the stability of the colour, and the effect of the presence of serum

Widespread serous membrane involvement by rheumatoid nodules.

Abstract: The characteristic cutaneous rheumatoid nodule, described fully by Collins (1937) and Bennett, Zeller, and Bauer (1940), is composed of a central area of fibrinoid necrosis around which develops an encircling corona of closely packed mesenchymal cells, the great majority of which appear to be fibroblasts (Collins, 1937), and external to this is a zone of inflammatory tissue. Lesions with similar histological features occur in the synovial membranes, and have been described involving serous membranes such as the pleura (Bennett et al., 1940; Baggenstoss and Rosenberg, 1943; Gruenwald, 1948), and Raven, Weber, and Price (1948), in whose case the peritoneum was also involved. Rheumatoid lesions have been known for some time to involve the pericardium (Baggenstoss and Rosenberg, 1944; Feiring, 1945; Clark and Bauer, 1947, 1948; Raven et al., 1948) the myocardium (Clark and Bauer, 1948), the ring and cusps of the mitral and aortic valves (Grzimek, 1932; Baggenstoss and Rosenberg, 1941; Bywaters, 1950; Sokoloff, 1953a), the aortic valve (Pirani and Bennett, 1951 ; Bauer, Clark, and Kfilka, 1951), and the tricuspid valve (Baggenstoss and Rosenberg, 1941; Gruenwald, 1948). Ellman (1947) and Ellman and Ball (1948) have described interstitial involvement of the lung in rheumatoid disease. It is therefore thought that the present case may be of interest as it illustrates the widespread clinical manifestations associated with rheumatoid lesions of the pericardium, myocardium, endocardium, all heart valves, sclera, visceral pleura, and dura mater. As far as can be ascertained involvement of the dura mater has not previously been recorded.

Malignant Cells in Cerebrospinal Fluid

Abstract: Ever since the beginning of the present century, the identification of the cells in cerebrospinal fluid (C.S.F.) has been recognized as an important diagnostic procedure. Unfortunately, it has always been found a difficult matter to make stained films of these cells, and in fact the preparation of Romanowsky-stained, air-dried films comparable with those of blood has been considered almost impossible (Reichmann, 1911). In spite of the introduction of several alternative techniques, none has found a permanent place in laboratory practice, and the preparation of a film is usually omitted. Presumably as a result of this difficulty, the number of reported cases in which tumour cells have been detected in C.S.F. is small. The fact that they can be found at all does not seem to be generally known. The purpose of this paper is to record seven cases of which six were subsequently proved to have meningeal spread of a malignant growth. (No post-mortem examination was permitted on the seventh.) These cases illustrate the importance of making stained films in cases of cerebrospinal pleocytosis, and a method for preparing such films is described.

Non-reactive tuberculosis.

Abstract: cases with leukaemia, myelosclerosis, and Hodgkin's disease have been isolated in the table. Forty-seven cases remain in the main group. Other cases are often quoted, but they will not be considered as they are unacceptable for the following reasons: Weichmann (1922), R6th (1929), Holzer (case 2, 1927), and Reiche quoted by BAlint (1925) give no histological description. The cases of Reilly and Bolin (1931) and Loeschcke (1932) were newborn infants and were probably cases of transplacental infection. The salient features of the 47 cases in the main group are summarized in Table II.

The use of a bile-aesculin medium and of Maxted's technique of Lancefield grouping in the identification of enterococci (group D streptococci).

Abstract: The name enterococcus, originally given by Thiercelin (1899) to a Gram-positive coccus of faecal origin growing in pairs and short chains, has been widely used for a group of morphologically similar streptococci found in the gut of man and other mammals, in dairy products, and in soil. The limits of this group, formerly rather vague, were satisfactorily defined with the application to it of Lancefield grouping methods by Sherman and his collaborators (Sherman, 1937; Sherman, Mauer, and Stark, 1937; Sherman, Stark, and Mauer, 1937; Sherman, Stark, and Yawger, 1937), and later by Shattock and Mattick (1943) and Shattock (1945 and 1949b). From this work it is apparent that the great majority, if not all, enterococci belong to Lancefield Group D, and that in fact this serological property should be taken as the basis of their classification. The group will then include the following species: Strep. faecalis, with its varieties, zymogenes, and liquefaciens, Strep. durans, and Strep. bovis. The degree of variation in the biological properties of Group D streptococci is well shown in Table I reproduced from Shattock (1949b ; see also Nyman, 1949). Table I covers the majority of important biological properties of the group, except bile tolerance, or the ability to grow in the presence of bile in concentrations up to 40% (Weissenbach, 1918), which is usually regarded as common to all species of enterococci, and aesculin hydrolysis. Attention was drawn to the value of aesculin hydrolysis in the identification of enterococci by Rochaix (1924), and the use of aesculin in bile media was regarded as the best single differential test by Meyer and Schonfeld (1926), who found that 61 of their 62 strains of enterococci hydrolysed aesculin, as compared with only six among 82 strains of Strep. viridans. Weatherall and Dible (1929), who obtained similar results, 60 of their 61 bile-tolerant, heat-resistant, mannitolfermenting diplococci hydrolysing aesculin, considered, however, that the hydrolysis of aesculin in a bile medium was \" very little more than an indicator\" of bile tolerance. This conclusion was probably responsible for the fact that the test has been little used. The work of Williams and Hirch (1950) suggests, however, that aesculin hydrolysis has an independent value in the classification of bile-tolerant cocci, as only 83.6% of their 140 strains of bile-tolerant streptococci hydrolysed aesculin. The present paper records an attempt to find a simple, rapid and accurate single procedure for the recognition of enterococci as a group, minor subdivisions of which are of little importance in the routine diagnostic work of a medical laboratory. Results of Lancefie!d D grouping were compared with those of tests for bile tolerance and aesculin hydrolysis, as well as heat resistance, on a number of freshly isolated and stock strains of streptococci.

An estimation of plasma vitamin A and the vitamin A absorption test.

Abstract: Following the adaptation of the Carr and Price (1926) colour reaction to the determination of the vitamin A content of the blood by McCoord and Luce-Clausen (1934), Chesney and McCoord (1934) reported impaired absorption of vitamin A from the intestine in coeliac disease, and devised the vitamin A absorption test as an index of the absorption of fat from the intestine. The chemical method was improved by Kimble (1939) and by May, Blackfan, McCreary, and Allen (1940), in each case a photoelectric colorimeter of the Evelyn (1936) type being employed in place of the earlier visual matching of the colours. The vitamin A absorption test has since been used by a number of workers in the United States, who confirmed the findings of low fasting plasma vitamin A levels and flat absorption curves in coeliac disease (Breese and McCoord, 1939; Clausen and McCoord, 1938; May et al., 1940; May, McCreary and Blackfan, 1942; Adlersberg and Sobotka, 1943a), in cystic fibrosis of the pancreas (Blackfan and May, 1938), in sprue and idiopathic steatorrhoea (Adlersberg and Sobotka, 1943b; Ingelfinger, 1943; Cayer, Ruffin and Perlzweig, 1945; Darby, Kaser, and Jones, 1947), in congenital atresia of the bile duct (May et al., 1940), and in colitis (Page and Bercovitz, 1943). It was soon found, however, that low plasma vitamin A levels and flat absorption curves may occur in conditions other than those characterized by steatorrhoea, particularly in chronic infectious diseases, e.g., in tuberculosis (Breese, Watkins, and McCoord, 1942), in giardiasis (Katsampes, McCoord, and Phillips, 1944), in catarrhal jaundice (Breese and McCoord, 1940), in cirrhosis of the liver (Ralli, Bauman, and Roberts, 1941), in malnutrition, inflammatory lesions, and cretinism (May and McCreary, 1941), and in several diseases of the skin (Ruch, Brunsting, and Osterberg, 1946; Peck, Glick, Sobotka, and Chargin, 1943). It is thus evident that the low plasma vitamin A level and flat absorption curve is not in itself diagnostic of steatorrhoea although both findings are invariably present when fat absorption is impaired. Ingelfinger (1943) considers that in assessing the low vitamin A level and flat absorption curve much is gained by a knowledge of the plasma carotene level, which is also low in steatorrhoea (Cayer et al., 1945). Our purpose in this communication is to present a method of plasma vitamin A estimation and to record data which we have found relevant to the performance and interpretation of the absorption test. Method

A review of the varieties of human haemoglobin in health and disease.

Abstract: THE OXYGEN-CARRYING FUNCTION OF HAEMOGLOBIN .. .. .. ELECTRONIC STRUCTURE OF HAEMOGLOBIN ANTD ITS DERIVATIVES .. .. Bond Type in Metal Complexes The Haem-Globin Bond GCENERAL PROPERTIES OF HAEMOGLOBIN AS A GLOBULAR PROTEIN .. The Properties of Haemoglobin in Solution Amino-acid Composition of Globin Immunological Properties of Haemoglobin The Solubility of Haemoglobin The Shape of the Haemoglobin Molecule Crystallography of Human Haemoglobin FOETAL HAEMOGLOBIN .. Crystallographic Differences Solubility Differences Differences in Amino-acid Composition The Immunological Specificity of Foetal Human Haemoglobin Alkali-denaturation of Adult and Foetal Haemoglobins ..

A micro-haematocrit for determining the packed cell volume and haemoglobin concentration on capillary blood.

Abstract: Gruneberg, 1942; Meyerstein, 1942; Smith, 1944; Harris, Gilding, and Smart, 1951 ; Sabine and Nickolai, 1952) and those open at both ends (Gram and Norgaard, 1923; Van Allen, 1925a and b; Guest, 1938 ; Kato, 1938 and 1940; Miller, 1939 Hamre, 1940; Benditt, Straube, and Humphreys, 1946; Shils, Sass, and Goldwater, 1952). All these tubes suffer from drawbacks either in design or method of use. A simpler capillary mnicro-haematocrit tube, open at both ends, heparinized and dried before use, and effectively

The use and limitations of filter-paper electrophoresis.

Abstract: In 1935 Tiselius, using the apparatus he had devised in 1930, demonstrated that human serum globulin fell into three reasonably defined groups based on their electrophoretic mobilities. For the next 15 years a steady flood of papers reported the fluctuations of albumin and the globulins in young and old, in health and disease, as disclosed by analyses carried out using Tiselius apparatus with and without the numerous minor modifications which were introduced. In spite of brilliant efforts at simplification it was evident that the method, because of its relative complexity, could never have widespread use in routine clinical chemistry. In 1950 three groups of workers, among them Tiselius himself, described methods which indicated that, using filter paper as a matrix, a relatively cheap and rapid method could be evolved of obtaining a rough analysis of the serum proteins akin to the analysis obtained by the classical technique. The method required no more than 0.5 ml. of serum, and though estimates of the total protein concentration would still have to be made by other means, it was conceivable that this simple technique would dispense with the tedious salt fractionation and subsequent analyses required heretofore to obtain the albumin/globulin ratio. The last four years have produced a spate of papers devoted to the description of modifications in methods of separation and analysis with numerous technical and mathematical tricks for improving the correlation of the results with the substantial body of data already obtained with the classical technique. With few notable exceptions scant attention was paid to the fundamental principles underlying the technique or to the points in which their application to it differed from the classical procedure. As paper succeeded paper it became increasingly evident that except within fairly wide limits the two methods were not quantitatively comparable. Even worse, because of lack of time spent on a consideration of the basic principles involved,

Specificity of Auto-antibodies in Acquired Haemolytic Anaemia

Abstract: Until recently it has been assumed that the autoantibodies found in acquired haemolytic anaemia are non-specific; that is to say they react with all types of human red cells and are unrelated to any known blood group antibodies. However, Weiner, Battey, Cleghorn, Marson, and Meynell (1953) have described a patient in whom the auto-antibody had the specificity anti-e, the patient's red cells having the genotype CDe/CDe: the same phenomenon had previously been suspected in a patient investigated by Dr. Ruth Sanger (personal communication). In the course of investigating a severe case of acquired haemolytic anaemia (Case 1), we noted that the patient's serum strongly agglutinated red cells of the genotypes CDe/CDe and cde/cde but not those of cDE /cDE. Further investigation showed that the patient's serum contained anti-C and anti-e, despite the fact that the patient's genotype was CDe/cde. This observation led to the re-investigation of nine other cases of acquired haemolytic anaemia whose warm auto-antibodies had been considered to be non-specific; the findings are described briefly in the present report.

Three Cases Illustrating the Presence of Argentaffin (Kultschitzky) Cells in the Human Gall-bladder

Abstract: Occasionally typical argentaffin (Kultschitzky) cell or so-called \"carcinoid \" tumours have been described in the human gall-bladder (Joel, 1929; Porter and Whelan, 1939; Bosse, 1943), although I have found no mention in textbooks of histology or pathology of the presence of such cells in the mucosa. There is, however, one case report (Kerr and Lendrum, 1936) of a papilloma of this organ covered with intestinal epithelium containing both Paneth and argentaffin cells. I propose to describe three cases in which the latter type was present.

A Note on the Serology of Sarcosporidiosis and Toxoplasmosis

Abstract: Serological methods for the diagnosis of toxo-plasmosis have received considerable attention since the first authentic report of a fatal infection in a human infant (Wolf, Cowen, and Paige, 1939). The complement fixation reaction (Warren and Sabin, 1942) and the dye test (Sabin and Feldman, 1948) are now recognized as the two best methods of detecting the disease. Serological studies in sarcosporidiosis have not, however, been given the same amount of attention, although this infection has a much longer history than that of toxo-plasmosis. For example, organisms designated as sarcosporidial in nature were described in the laryngeal muscle cells of a human patient (Baraban and Saint Remy) as long ago as 1894. Similar aggregations have since been described in isolated human patients in the muscles of tongue, biceps, heart, etc., and these have usually been given the specific name of Sarcocystis lindemanni (Rivolta). The discovery that chronic toxoplasmic infections are often associated with the formation of \" pseudocysts \" containing large numbers of organisms not only in nervous tissue but in skeletal and heart muscle adds to the suspicion that some reported cases of sarcosporidiosis in man and animals might in fact be cases of toxo-plasmosis. Different investigators give different dimensions for these organisms. \" Sarcosporidial spores \" described in human infections range from 8 to 9 ,u long (Baraban and Saint Remy, 1894) to 4.25 p long (Darling, 1909, 1919). While it seems unlikely that organisms regularly of the first dimensions are Toxoplasma, suspicion arises in the case of smaller organisms. The issue is complicated by the fact that in some of these reports the \" septa\" and enclosing membrane associated with the more typical sarcosporidial cysts could not be demonstrated. Kean and Grocott (1945) concluded that in the absence of inoculation and serological studies the absolute identification of these two parasites is impossible. In October, 1951, Muhlpfordt used the Sabin-Feldman dye test in an endeavour to differentiate between infections of Toxoplasma and Sarcocystis. \" Miescher's tubes \" (sarcosporidial cysts) recovered from gullet muscles of sheep or goats were inoculated per os or intraperitoneally into guinea-pigs, rats, and hamsters, and the serum of these animals was tested by the dye test against toxoplasms. In all cases, preliminary tests made before the inoculation with sarcosporidia produced negative results, but after the infection was allowed to establish itself the animals reacted positively , thus suggesting that antibodies reacting with the toxoplasms were …

The Estimation of Heparin and Similar Substances in Human Blood and Tissues Using a Combined Biological and Colorimetric Method with Paper Electrophoretic Studies

Abstract: Heparin and other acid mucopolysaccharides were extracted from various tissues by a modification of the method previously described (Bassiouni, 1953). The dye mucopolysaccharides complex obtained was resolved into its components with formamide: the acid mucopolysaccharides were precipitated with methyl alcohol in the presence of electrolyte (sodium chloride). A solution of the purified polysaccharide precipitate was bio-assayed for active heparin, and the remainder was either submitted to electrophoresis on paper

Acute haemorrhagic leuco-encephalitis.

Abstract: Itwas in1940thatWestonHurst(1941) encounteredtwo cases offatalencephalitis whichhe regarded on clinical andpathological grounds as representing a previously unidentified entity to whichhe gave thename ofacutehaemorrhagic leuco-encephalitis. To quoteHurst'soriginal description, thedisease was "an acute'cerebral condition, virtually localized asfarasthecerebrum was concerned tothewhite matter, anddeveloping more or lessabruptly in apparently normal individuals. Perivascular necroses,perivascular andfocaldemyelination, haemorrhages, oedema, andcellular infiltration were thechiefcomponents ofthepathological picture." Inthe12yearssinceHurst's paperappeared remarkably fewsimilar caseshavebeenreported, andinfactonlyfivefurther examples havebeen described underthesame name. Thesehavebeen recorded byHensonandRussell (1942), Shallard andLatham(1945), MacArdle, van Bogaert, and Lhermitte (1949, two cases) andby Greenfield (1950). Itso happensthatthreecharacteristic examples ofthedisease haveoccurred inthe neurosurgical department ofSt.George's Hospital over a two-year period, anditseems probable that thecondition isnotsuchan extremerarity asthe smallnumberofrecorded caseswouldindicate. Theobject ofthepresent communication istoput on record thesethreefurther cases(bringing the numberrecorded up to10)andtodrawtheattentionofgeneral pathologists totheexistence ofthis variety ofencephalitis. Thedescriptions ofthe disease whichfollow arebased on thethree new casestogether withthesevenpreviously reported. Moredetailed summaries ofthethree cases are included inan appendix tothis paper. Clinical Features

The Appearance and Significance of Tissue Mast Cells in Human Bone Marrow

Abstract: Despite an increasingly vast literature concerning the examination of bone marrow, scant attention has been paid to the occurrence of tissue mast cells in human marrow or to their significance when present. Only recently have mast cells been described in the marrow of a small number of cases by Rohr (1949), Leitner (1948, 1949), Bremy (1950), Tischendorf and Hartmann (1950), Fadem (1951), Koszewski (1952), and Hayhoe (1953). Generally the patients had a severe marrow disturbance, which was frequently of the hypoor aplastic variety, and, in the opinion of both Undritz (1946a and b) and Bremy (1950), the appearance of mast cells in the marrow indicates severe marrow depression and is of diagnostic and prognostic significance. However, Williams (1952), using marrow biopsy particle smears, was able to demonstrate mast cells in as many as 56 (17 %) of 325 marrows. Fixed tissue sections of routine marrow aspirates were used in the present work, the object of which was to observe the frequency with which tissue mast cells occur in human marrow and to try to assess their significance.

Sertoli and Leydig cells in relation to ovarian tumours.

Abstract: The group of tumours termed arrhenoblastomas was separated from the general body of ovarian tumours by Meyer (1915, 1930, 1931). Barzilai (1949) defines the arrhenoblastoma, in conformity with Meyer's views, as \" an ovarian tumour the cytoarchitecture of which almost exacly duplicates that seen in the different stages of male gonadogenesis.\" Thus, although many of these tumours have a masculinizing or defeminizing effect on the host this is not essential for the diagnosis. Descriptions of about 122 of these tumours have been published (Javert and Finn, 1951). Three types of tumour pattern are recognized: (1) a highly differentiated pattern composed of well-developed tubules lined by cubical or columnar cells with basal nuclei, closely resembling Sertoli cells and the late developmental stages of gonadogenesis, as seen in the undescended testis ; (2) one in which the tumour looks like a fibromatous or sarcomatous neoplasm; and (3) a pattern intermediate between these two extremes. Barzilai (1949) and Novak (1952) have drawn attention to the large polyhedral eosinophilic cells which frequently lie in the stroma of the tumours and resemble Leydig cells. The work of Sternberg (1949) and of Teilum (1949) allows the separation from this heterogeneous group of two distinct entities (a) \" Leydig\" cell tumours and (b) \" Sertoli \" cell tumours ; a third form of tumour also exists in which both \" Sertoli \" cells and \" Leydig\" cells occur. In this paper it is proposed to describe and discuss two tumours, which are morphologically arrhenoblastomas, in the light of these newer concepts, and also a case of bilateral Leydig cell tumour, or hyperplasia, with masculinization.

Combined agar diffusion and replica plating techniques in the study of antibacterial substances.

Abstract: Much interest has recently been aroused in the field of antibiotics by two distinct but related problems. The first concerns the possibility that in certain types of infection, notably subacute bacterial endocarditis, the bactericidal action of a drug is of greater therapeutic significance than the inhibitory effect (Hunter 1950; Robbins and Tompsett, 1951 ; Cates, Christie, and Garrod, 1951). The second problem is the action of antibiotics used in combination. In this respect the findings of many workers indicate that different mixtures may show augmented or diminished bactericidal effect, and the result may be influenced by the species or strain of organism tested (Jawetz, Gunnison, Bruff, and Coleman 1952; Bliss, Warth, and Long, 1952). Any investigation of these two problems requires a method applicable to routine use for the assessment of the bactericidal action of antibiotics. None of the conventional methods used for these purposes is suitable for large-scale application, as they depend on some form of viable counting and are very laborious. Even the more convenient turbidimetric technique described by Burnell and Kirby (1951) is too elaborate for general use. Moreover, a disadvantage common to all these methods is that an appreciable amount of drug is inevitably transferred with any sample to the new medium used to measure the bactericidal action. If the proportion of surviving bacteria is low it may be necessary either to limit the range of concentration of the drug under study, or to destroy it in some way which does not affect the bacteria. This is not always possible or convenient. A method of transfer which carries over little more than the amount of drug adhering to individual organisms is therefore needed. The method described here, which is an application of the technique of \" replica plating\" described by Lederberg and Lederberg (1952) and used by them for studying problems of bacterial genetics, is believed to fulfil these requirements. It involves no significant transfer of drug from one medium to another, is simple enough for routine use, and may be applied as a continuation of a form of \" sensitivity test \" in common use. It consists essentially of the transfer of bacteria from one nutrient agar surface to another without disturbing their spatial relationships. The transferring agent is the pile of a piece of sterile velvet covering a flat surface. The success of the technique depends on the fact that moisture is absorbed by the pile and no lateral smearing of the impression occurs.

The Classification of Anaerobic Cocci and their Isolation in Normal Human Beings and Pathological Processes

Abstract: In a previous paper (Hare, Wildy, Billett, and Twort, 1952) a scheme employing fermentation of carbohydrates and organic acids was put forward for the classification into six groups of the anaerobic cocci derived from human beings. There is at present extremely little information as to the occurrence of anaerobic cocci on the mucous and skin surfaces of normal human beings. This, together with a study of strains from a number of infective processes, forms the subject of this paper.

Haemorrhagic Diathesis Due to Deficiency of Factor VII

Abstract: Of recent yearsithasbecomeapparent that Quick's one-stage prothrombin time, although a mostuseful routine laboratory test, doesnot specifically measure plasmaprothrombin. Quickhimself (1947) recognized thatdeficiency ofhislabile factor (Factor V)caused lengthening of theprothrombin time,and Owren(1947) reported thefirst caseofcongenital deficiency of Factor V causing a haemorrhagic state whichhe termed "parahaemophilia." Itisnowknownthataprolonged prothrombin timemayresult fromlackofanother factor which normally develops inserumduringcoagulation, andhasbeennamedbyvarious workers "serum prothrombin conversion accelerator (Alexander), "convertin " (Owren), andFactorVII(Koller, Loeliger, andDuckert, 1951). Onlytwo undoubtedcasesof congenital deficiency of thisfactorhaveso farbeen described, thefirst by Alexander, Goldstein, Landwehr, andCook(1951), and another by Owren(1952). Thefollowing caseappears tobe thethird example. CaseReport

The Estimation of Total Nitrogen Using the Conway Micro-diffusion Cell

Abstract: The use of the microdiffusion unit for the determination of blood urea has been described by Conway and O'Malley (1942). Recovery of urea added to blood when estin;ated by this method is precisely quantitative as compared with a 98 to 99% recovery given by the aeration method (Kay and Sheehan, 1934). The method has been found very useful for routine laboratory determinations, especially when dealing with large numbers of samples. Conway (1939a) has also described a method for determining total nitrogen, using the diffusion cell to estimate the ammonia after digestion by the usual Kjeldahl technique. A modification of the method so that the same reagents as those used for blood urea estimation can be employed is described. The method is of considerable value, especially when only small amounts of sample are available or large numbers of estimations have to be performed, since distillation is avoided and all titrations may be carried out together, if necessary on the day after setting up the units.

A Rapid Method of Identifying Particular Barbiturate Derivatives in Small Samples of Blood by Paper Chromatography

Abstract: The quantitative estimation of barbiturates in blood by ultra-violet spectrophotometry (Goldbaum, 1948; Walker, Fisher, and McHugh, 1948; Born, 1949; Lous, 1950; Wright and Johns, 1953) is of limited value unless the particular barbiturate derivative is known or can be identified. Goldbaum (1952) described an ultra-violet spectrophotometric technique by which he claimed to make such identification, but many workers with experience in this field will prefer to rely on more direct methods of qualitative analysis. For general purposes the micro-crystalline method of Turfitt (1948) is suitable if urine or gastric contents are available, though even then it is always best to confirm the identification by extraction and mixed melting point determination. Cases of barbiturate poisoning frequently occur, however, where such material is not available for analysis, or if available, does not contain adequate amounts of barbiturate, as in poisoning by short-acting derivatives. It is in these cases that a qualitative method applicable to small blood samples is of particular value. Paper chromatography seems suitable for this purpose.

A Family Illustrating the Double Inheritance of the Sickle Cell Trait and of Mediterranean Anaemia

Abstract: an exact explanation of hitherto imperfectly underthat the red cells stood phenomena. This is well illustrated by the separate haemoglob recent advances made in the understanding of sickle haemoglobin III or cell anaemia and the sickle cell trait. In 1949, 1951) and haemogl Pauling, Itano Sir ger, and Wells demonstrated that are thus five varic the carbon monoxide derivative of the haemoglobin normal adult or A, derived from the red cells of sickle cell anaemia S, and haemoglobim possessed a different mobility from normal haemoof Health, 1953) * globin. They showed that the red cells of sickle By combining th cell anaemia contained almost entirely this \" sickle \" phoresis and by q haemoglobin, whilst the red cells of sickle trait (Singer) the red ce carriers contained about 50% sickle haemoglobin usually found to co and 50% normal haemoglobin. In 1950, Perutz 1000% and foetal has and Mitchison demonstrated that reduced sickle red cells of the sick] haemoglobin was extremely insoluble and the globin 5 to 450% ani crystals formed were similar to red cells that had globin. The red ce: undergone \" sickling.\" Thus sickle cell haemoglobin form of Mediterrai joined normal haemoglobin and foetal haemoglobin adult haemoglobin

The thromboplastic activity of Russell's viper venom and its relationship to factor VII.

Abstract: as a powerful local haemostatic agent in the treatment of haemophilia. Fullerton (1940) substituted Russell's viper venom for rabbit-brain thromboplastin in his modification of Quick's one-stage prothrombin time, since viper venom could be obtained in a more convenient and more stable form than brain extract It soon became apparent, however, that the two methods gave widely different results. Wilson (1947) and many other workers found that in cases treated with dicoumarol much higher plasma prothrombin values were obtained with venom than with brain thromboplastin, and, in fact, Fullerton's modification could be dangerous, since it gave inadequate warning of prothrombin values low enough to cause haemorrhage. It has been shown by Macfarlane, Trevan, and Attwood (1941) that a lipoid substance in the plasma is necessary for the action of viper venom, and an excess of lipoid, such as occurs after a fatty meal, shortens the prothrornbin time with venom but not with brain thromboplastin (Fullerton and Anastasopoulos, 1949). A further source of the difference in action of the two thromboplastins became apparent when it