[Demonstration of antibodies to salmonella-R-antigens in sera of patients suffering from salmonelloses (author's transl)]. The extent of hemagglutinating antibody titers against Salmonella R-lipopolysaccharides in 189 sera of 119 human beings suffering from various salmonelloses was estimated. When compared to serum specimens of 137 healthy persons a significant rise in the number of Ra- and Rb2-titer containing sera was observed. It was further found that titers directed against chemotypes Ra, Rb2 and Rc were significantly enhanced in height when compared to the control group. Sera of healthy persons and patients did not show significant differences in Rd1-, Rd2- and Re-titers, respecively. The results indicate that the innermost part of the LPS core in Salmonella S-forms is considerably less (or even none) immunogenic in respect to antibody formation. R-titers in sera from patients suffering from salmonelloses as well as from healthy persons were predominantly directed against Ra-, Rb2- or Rc-LPS structures; the Ra-titer being the most frequent one in both groups. More detailed analyses on sera from patients suffering from salmonelloses revealed a somewhat lower content of R-titers in the early stage of infection. Furthermore in typhoid fever changes in the height of Ra-titers paralled the usual course of O-titers and in gastroenteritic infections the course of illness, respectively. The immunogenicity of the LPS core, however, was less expressed than that of the O-polysaccharides as revealed from a comparison in extent and height of R- and O-titers. Frequently the salmonelloses sera showed titers against several R-antigens. In such cases Ra-titers mostly occurred either together with Rb2-titers alone or concomitantly with both Rb2- and Rc-titers. In absorption experiments it could be demonstrated that two types of antibodies had been formed, those directed against terminal sugar residues in R-lipopolysaccharides and others with specificties against internal partial structures of the core.


The unfortunate role of precedent in bacteriology. II. Unrelated serological specificities of the polysaccharides from S. typhi and S. gallinarum sharing factors 9.12. Tube and agar-gel precipitations of antigens isolated from S. typhi and S. gallinarum against homologous and heterologous rabbit antisera showed that the polysaccharides of these Salmonellae, sharing factors 9.12 do not cross-react. Thus, the serological analyses prove that sugars can not be related to factors in the Kauffmann-White scheme. The cross-reacting antigens from these Salmonellae of serogroup D are the free proteins and the proteinic moieties of their somatic antigens.


[Air pollution by motor vehicles along the freeways]. Total mass particulates and benzopyrenes have been determined to evaluate the specific pollution by vehicles on a section (6 km) of a highway. The traffic density generated an increase of 30% mass particulates in nighttime and 44% during daylight. Concentrations of benzopyrenes in air have doubled compared to the period before traffic.


Sh. dysenteriae serotypes2,4,8-immunochemistry and phage receptor activity. Among three analyzed serotypes of Shigella dysenteriae, namely, the serotypes 2,4 and 8, the serotype 2 proved to be a strong immunogen in rabbits, inducing anti-polysaccharide antibodies as well as antiprotein antibodies in all the animals. In contrast, the serotypes 4 and 8 were weak immunogens and among the rabbits some have synthesized only anti-proteins while others had antibodies against the somatic conjugate. Aside from the somatic antigens, large amounts of proteins were isolated from all the strains; however, the numerous determinants of these proteins were proven with the help of a serum to proteins from Sh. sonnei. The polysaccharides were specific for the serotype. The sensitivity of Sh. dysenteriae strains to phage P1 and the phage receptor actigity of different bacterial extracts were examined. By using the phage receptor neutralization test, it was possible to demonstrate that the receptor substance is a common component present in the lipopolysaccharide. The nature of neutralization has been discussed.


[Measurement of exhaust gases of cars in the neighbourhood of roads (author's transl)]. Air samples were collected in plastic bags simultaneously at various measuring points in the close range of streets. When examining the various bag materials, Teflon bags showed the smallest deviations in direct analyses and in analyses of up to two hours after the drawing of samples. The following methods were used for the analysis of the air samples collected in the bags: coulometry for CO and SO2, chemiluminescence for NO/NO2, chromotropic acid for CH2O and flame ionization for hydrocarbon. The various components were measured close to a highway and near streets in residential and business areas.


[Detection of alkylating air pollutants (author's transl)]. Occupational toxicological observations made during the last few years have shown that the respiratory intake of certain alkylating compounds entails a major carcinogenic risk even at concentrations below 1 ppm. Such inhalation carcinogens can also stem from photochemical and spontaneous chemical reactions in the atmosphere and not merely from direct emissions. For the examination of air samples for alkylating activities simple colorimetric test methods are available. Some of them must be considerably modified and matched to the reaction of the various alkylating compounds. In this manner colour reactions of, in part, high selectivity are obtained: thus, for example, carcinogenic alpha halogen ether can be detected in concentrations of 10 ng/ml test solution. By the use of sensitive colorimetric methods of analysis it was possible to fractionate the alkylating activity found in the exhaust gases of automobiles. It is now possible to enrich certain compounds, which react positively to the colour test, and to purify them for further chemical characterization.


[Several developmental disorders in children with schizophrenia]. The authors discusses some unclear and insufficiently studied problems related to early infantile autism, regress of development and underdevelopment of schizophrenic children. The basis of early infantile autism is most likely a peculiar disturbance of development due to constitutional, organic and psychogenic factors. In most of the cases this syndrome is connected with the schizophrenic process. The majority of psychiatrists in the Soviet Union consider lowering to a more early level of development as a regress of development in child schizophrenia symptoms. The clinical picture of retarded development in childhood schizophrenia depends upon the age of the onset of the disease and the degree of progressiveness of the process. Depending upon these criteria it is possible to distinguish retarded development, resembling oligophrenia and phenomena of psychophysical infantilism.


The haematopoietic supraneural organ of adult, sexually immature river lampreys (Lampetra fluviatilis [L.] Gray) with particular reference to azurophil leucocytes. The haematopoietic tissue in the supraneural organ of the freshwater river lamprey (Lampetra fluviatilis L. Gray) was studied in sexually immature animals. Besides erythro- and granulopoietic elements, macrophages, reticular cells, fibroblasts and glycogen-rich fat cells were seen. Developing granulocytes of the lamprey contain one type of azurophil granules originating from small cytoplasmic (Golgi) vesicles. The lamprey's azurophil granulocytes seem to be homologous with those of fishes. However, the granulocytes of fishes, studied thus far, show granules with only one type of inclusion, whereas in lamprey the granulocyte inclusions are variable in size and shape. Thus, lamprey granulocytes are, in this respect, reminiscent of similar cells of higher vertebrates. The PAS and alkaline phosphatase reactions, common markers of vertebrate neutrophil leucocytes, are very weak in the haematopoietic tissue granulocytes of the lamprey, and intense in the blood cells of the same animal. Lamprey granulocytes, similarly to the granulocytes of Chondrostei and Elasmobranchiata, do not stain with peroxidase, naphthol-AS-D-chloroacetate esterase and sudan black B. The haematopoietic tissue contains a relatively high number of degenerated granulocytes.


The effect of hypothyroidism on the female genital tract of gerbils (Meriones hurrianae jerdon). The effects of hypothyroidism on the female genital tract of gerbils have been studied. Hypothyroidism was produced by (a) surgical ablation, and (b) pharmacological suppression of the gland. Hypothyroidism resulted in atrophic ovaries. Follivular development was severely arrested, with most of the follicles showing atresia. Distinct effects were produced upon the uterine physiology. Uterine regression was conspicuous in thyroidectomized females. Hypothyroidism resulted in a decreased RNA, protein, sialic acid and glycogen concentration of the uterus. Vaginal RNA, protein and sialic acid contents were low after thyroidectomy. The vaginal cytology showed a constant pattern of its cells, i.e. dioestrous. L-thyroxine treatment restores the biochemical changes of uterus and vagina to subnormal levels in thyroidectomized animals. It is concluded that hypothyroidism affected the weight, cytology and biochemistry of the female genital tract of gerbils.


Effects of small doses of L-thyroxine and triiodo-L-thyronine on pituitary LH content of thyroidectomized rats. Pituitary LH was studied by means of a specific radioimmunoassay (RIA) in male rats at different time intervals after thyroidectomy (T), and in rats which were T at least 30 days before and were then treated with different doses of L-thyroxine or triiodo-L-thyronine. A decrease in the pituitary LH of the T animals, with respect to the intact age-paired controls, was demonstrable from 13 days after the operation, when total pituitary LH content was taken into consideration, or from 5 days after T, when the LH concentration (mug/mg pituitary) was considered. Doses of thyroid hormones lower than the daily maintenance dose for the rat produced very little effect on the pituitary LH levels of T animals. However, a single dose of 1.75 mug of T4 or 0.2 mug of T3 (doses approximately equivalent to the T4 and T3 maintenance dose for T rats) induced such a rapid and intense increase in the pituitary LH content that it no longer differed from that of the intact age-paired controls by 12 hours. Surprisingly, 5.0 mug of T4 and 1.0 mug of T3 did not produce any increase in the pituitary LH content of T rats.


Diurnal variations of plasma and pituitary thyrotrophin in adult male and female rats. Male and female rats fed a low iodine diet for 20 days were used to study the diurnal variations in resting levels of plasma and pituitary TSH concentration using a highly sensitive radioimmunoassay. Sex differences in the fluctuations in plasma TSH levels and in amount of TSH in the pituitary gland were observed. The daily fluctuations of plasma TSH were characterized by two peaks that occurred in males at 6 a.m. and at 3 p.m. while in females the peaks were delayed until 9 a.m. and 7:30 p.m. Moreover, in the females the morning and the afternoon peaks were of the same intensity while in the males the afternoon peak that occurred just before the onset of darkness was much greater than the morning peak. There was a fall in TSH content of the pituitary in the male rats at 6 a.m. and also in the afternoon just before the onset of darkness. Thus, the diurnal variations in the plasma and pituitary TSH levels were related in male rats. In the females, however, the pituitary TSH concentration did not reflect the changes observed in the plasma TSH levels. The level of plasma PBI did not appear to be responsible for the fluctuations in plasma TSH concentration. It is suggested that the main mechanism for the control of the circadian rhythm of TSH might be related to a high activity at night.


Oat-cell bronchial carcinoma. Identification of cells in pleural fluid. An analysis is made of the accuracy of diagnosing oat-cell (small cell) carcinoma of the lung from the cytology of pleural fluid. All the material over a 22-year period has been reviewed. Air-dried smears were used, stained with May-Grünwald-Giemsa. Of 91 cases in which oat-cells were specified in the written report, 90 were either proved histologically to have oat-cell or anaplastic bronchial carcinoma, or else had clinical findings consistent with that diagnosis and were registered as having bronchial carcinoma at death. There was one false positive, and no false suspicious reports. Cytologic diagnosis of this tumor type has, therefore, been no less reliable than would be expected from a histologist examining a surgical biopsy of infiltrated tissue. Of 49 consecutive cases histologically proved to have oat-cell carcinoma, and with pleural effusions examined in this laboratory, 21 showed no malignant cells (43 per cent false negative). Diagnostic features and causes of error are discussed, as well as the advantage of air-dried smears over the standard Papanicolaou method for the detection of this cell type.


Immunopathogenic mechanisms in tissue damage. Our knowledge on immunopathogenic damage is derived from multiple different sources: experiments in vitro, experiments in vivo, clinical observations, and observations using material from multiple different animal species including man. If we want to analyse an immunological process suspected of causing tissue damage we have critically to evaluate the mechanism of immunologically specific initiating factors and the mechanisms of non-specific mediating factors. It is necessary to have clear information on the identity of the antigen, the location of the antigen, the identity of antibody, the location of antibody and similar information on specifically reactive lymphocytes. With this information in mind it is possible to hypothesize upon mechanisms, which can be responsible for the immunologically specific initiation of immuno-pathogenic damage. The non-specific mediating systems, which are the actual inducers of functional or structural tissue damage, are triggered by the specific, initial immune reaction. These systems comprise for instance complement, coagulation, granulocyte and macrophage lysozomes and function, chemotaxis, the mast cell system, phagocytosis etc. The overall picture of non-specific mediation of tissue damage is extremely complex, since it really makes it necessary to investigate and describe not only these systems but also their co-existence and interaction. Knowledge of immunopathogenic mechanisms in clinical disorders in man is still resting mainly upon analogies and assumptions, although progress is taking place. It is essential to ask the right questions and critically to evaluate the answers. Thereby the understanding of immunopathogenesis in diabetes mellitus and other clinical disorders shall gradually improve.


Insulin--anti-insulin complexes.

Insulin--anti-insulin complexes. When guinea pig antibodies (ab) bind insulin (ag), they can make complexes of different sizes. We propose the following model: In ab excess: (see article) Intermediate: (see article) In ag excess: (see article). An insulin molecule acts as a bivalent antigen, although more than two different antigenic determinants may be present. In vivo the large C II type disappears more rapidly from the blood than does the C I. The C II binds to complement factor C1q, whilst C I and C III do not. In sera from insulin treated patients we found C I and C III. The lack of lattice formation, due to the bivalency, may explain the difficulty in obtaining precipitation. The different complexes may influence calculations of antibody concentrations and affinity constants of the binding sites. The in vivo effects and possible clinical effects of antibodies to insulin may depend on the type of complex formed. Possibly, prevailing C II formation tends to cause large insulin requirements, although C II may seldomly be detected in the blood, because of rappid trapping. The immune complexes could affect the progression of angiopathy a) by interfering with insulin metabolism and control of diabetes, and b) by complement activation (mainly C II) and trapping in the vascular bed.


Sudanophilia in exfoliated urothelial cells. Cytoplasmic lipid globules colorable by Oil Red O of varying numbers, size and random distribution were encountered in the majority of normal, abnormal and malignant urothelial cells of the urinary sediments fixed by formaldehyde. Small cytoplasmic lipid globules and the diffuse sudanophilia of the finely dispersed lipids which paralleled the cyanophilia in the homologous cells were considered indicators of the functional status of well preserved urothelial cells; medium and large lipid globules were a sign of advanced necrobiosis. Demonstration of cytoplasmic lipids was not found a reliable discriminatory guide in the differential diagnosis of malignancy of the urothelium.


HLA factors and immune function.

HLA factors and immune function. The HLA system is the major histocompatibility system (MHS) in man and contains a variety of closely linked genes, which control a large number of alloantigens, some complement components, and certain immune responses. The alloantigens are of three different kinds: (i) so-called "classical" or serologically detectable (SD) antigens which are present on all nucleated cells and controlled by genes of at least three different loci; (ii) MLC (= mixed lymphocyte culture = LD = lymphocyte dependent) antigens which are as yet only recognizable by in vitro culture assays; (iii) Ia (= immune-associated) antigens which (like MLC antigens) primarily are present on B lymphocytes and which can be detected serologically. Our knowledge of immune-response (Ir) determinants in man is as yet rather limited but by inference from studies of MHS in mice, guinea-pigs, and rhesus monkeys it is almost certain that HLA contains such genes. Immune response genes control in a dominant way the ability to respond specifically with IgG antibody production and cell-mediated immunity to certain specific antigens. Ir genes are closely linked to the MLC and Ia genes. They seem to be particularly important for immune responses involving T lymphocyte function (including T-B cell cooperation), and it seems likely that they code for "antibody-like", carrier-specific immune receptors on T cells. Evidence that HLA contains Ir determinants derives from the observations (i) that a variety of "immunopathic" disorders are associated with HLA factors; (ii) that ragweed hayfever seems to be inherited with certain HLA types within families; and (iii) that the occurrence of anti-adrenal antibodies is associated with HLA in Addison's disease. Autoimmune thyreoiditis is controlled by the MHS in mice and chickens, and in analogy, Graves' disease is associated with HLA in man.


Application of alcian blue in the electron microscopic study of mouse and human cerebral cortex nerve cells. Alcian blue is a cationic dye which has been used in the histochemical field for the demonstration of polyanions especially carboxylated and sulphated. The results obtained in neurons when this dye was applied to human and mouse cerebral cortex and studied with the electron microscope are the object of the present report. The CNS of normal adult mice was fixed by vascular perfusion with 2% glutaraldehyde-0.1 M sodium cacodylate-0.1 M sucrose at pH = 6.8 followed by the same fixative with the addition of 0.5% alcian blue. After perfusion, brain cortex was taken out, sectioned into small blocks and immersed in a fresh similar mixture and subsequently in OSO4. Blocks were dehydrated and embedded in araldite. Ultrathin sections were doubly stained with uranyl and lead salts. Human brain cortex taken from patients with cerebral edema was fixed by immersion with 6.5% glutaraldehyde-0.1 M sodium phosphate, pH = 7.4 followed by embedding in warm agar and sectioning in slices of 30 mum thickness which were impregnated by immersion in a mixture of 1% alcian blue-acetate buffer-3% glutaraldehyde at pH = 3.5 for 9 to 15 h at  4 degrees C and subsequently immersed in 1% buffered OSO4-0.1 M sucrose, pH = 7.4 for 2 h at 4 degrees S. Sections were dehydrated and embedded in araldite. Ultrathin sections were doubly stained by uranyl and lead salts. We have denominated the complete procedure in both instances GABOUL technique. The submicroscopic study of both tissues, at nerve cells, revealed the presence of an electron dense homogeneous substance thoroughly dispersed at the hyaloplasmic matrix of perikarya, processes and even synaptic endings. This substance was more evident around free and attached ribosomes, GOLGI apparatus, complex vesicles, dense bodies, microtubules, subsurface cisternae and synaptic vesicles. Canaliculi of endoplasmic reticulum and even the perinuclear cistern also showed a moderate content. It is suggested that this electron dense substance, being alcianophilic, has a polyanionic character and thus may partially correspond to acid polysaccharides since these compounds have already been previously confirmed in such neurons by biochemical and light microscope histochemical techniques.


Improved periodic acid-silver methenamine staining procedure for electron microscopy. Mucosubstances can be stained for electron micrsocopy with silver methenamine after periodic acid oxidation in ultrathin Epon sections mounted on nickel grids. The grids are transferred from a solution to another during the staining procedure conveniently in a container made of polyvinyl chloride plastic. This staining procedure is easy to perform and gives reproducible results. It is especially suitable for simultaneous handling of a large number of grids.


[Therapy evaluation in rheumatology with an electronic data processing system]. A system for the evaluation of therapeutical effects in rheumatic diseases is presented. This system permits a calculation of two indices: i.e. rheuma-number and "Masszahl" (= only measured values such as range of motion in degrees) which lend themselves to statistical treatment. The result of standardized physical joint examination of patients is recorded on two mark-pages, specially developed for documentation of joint status. The data recorded on the two-mark-pages are then entered into the computer by means of a mark-page-reader. On the basis of these data, the computer, upon the basis of a program specially developed for this purpose, automatically calculates the corresponding amount of negative-points, which parallels the severity of the joint changes, i.e. the rheuma-number ("Rheumazahl"). Separately listed at the end of the status-outprint is the "Masszahl", which is part of the rheuma-number and comprises the measured values (for example, the range of motion measured in degrees). The practicability of this system is demonstrated on the basis of the results of a double-blind-comparison of two antirheumatic-corticosteroid-combinations and an open study with immuno-suppressives in patinets with rheumatoid arthritis.


General health screening of four-year-olds in a Swedish county. IV. An analysis of the effectiveness of the psychological examination program. Since 1969, 4-year-olds in the County of Uppsala have been offered extensive health screening at the Child Health Centres. In 1969 and 1970 a total of 3810 children underwent the screening procedure. The psychological screening instrument consisted of a questionnaire (to be answered by the parents), an interview of the parents, a psychomotor examination and an observation of the child's behaviour. 156 (4.1%) 4-year-olds were referred to a specialist team for investigation and decision concerning treatment. In the present study the effectiveness of the psychological screening instrument was assessed by its sensitivity, specificity and predictive value. The sensitivity was found to amount to a maximum of 0.73. The specificity was estimated to be 0.98-0.99, and the positive predictive value to 0.67. The authors consider the effectiveness to be sufficiently high to motivate continuation of the screening procedure for detection of psychological problems in pre-school age. Efforts should be made, however, to further improve the effectiveness.


Nerve fibers in the human organ of Corti. In the human cochlea, the following characteristics of nerve fiber arrangements were observed. 1. Where there was only the basilar fiber in the tunnel of Corti, the internal and tunnel spiral bundle were not observed. In such cases, the number of nerve fibers in the external spiral bundle was small. 2. In the tunnel of Corti, the basilar fibers (afferent) took a course at the base and crossing the tunnel slanting to the basal end. 3. The medial fibers were present with the internal spiral bundle and tunnel spiral bundle. These fibers are possibly efferent. 4. The external spiral bundle increases its fiber count when internal and tunnel spiral bundles were present in the organ of Corti. 5. The external spiral bundle was composed of the basilar and medial fibers. The former were less and the latter much more numerous in the bundle. 6. 85-90% of the afferent fibers went to the internal hair cells, while the remainder went to the external hair cells. 7. The nerve fibers were found in the organ of Corti lateral to the external spiral bundles. The fibers were running in the Hensen's and Claudius' cell area. 8. In some cases medial fibers showed a torpedo-like deformity in the tunnel of Corti. When this was observed, it was found distributed throughout the entire cochlea. Hearing disorders due to this pathology of the efferent fiber was suggested.


Selective loss of blood group antigens during wound healing. Frozen sections of healing wounds of the oral mucosa of 3 rhesus monkeys were examined by a double layer immunofluorescent technic for the presence of blood B-like antigens. Antigen activity was present in normal epithelium and in the epithelial outgrowth into the wounds but was absent from the outgrowth following treatment with lipid solvents. This finding suggests that glycoprotein but not glycolipid associated antigen is lost from the surface of epithelial cells during wound healing, a pattern of selective loss which may reflect a difference between regenerative and neoplastic lesions.


Fibrinoid necrosis of the epithelial cells of the skin. It has been shown in previous studies that fibrinoid necrosis not only occurs in connective tissue and in vessel walls, but can also be observed in the liver cells under special conditions and can also be provoked experimentally. It was observed in the present study that certain dermatoses (cases of the herpes group, erythema multiforme, drug eruptions, Lyell's toxic epidermal necrolysis, pityriasis lichenoides acuta, and skin affected by UV-rays) were associated with "eosinophilic necrosis" in the epithelial cells which morphologically corresponded to the fibrinoid necrosis of the connective tissue and of the liver cells described previously. Besides toxic, infectious and septic conditions, circulatory disturbances (hypoxia, anoxia) appear to have a special significance. Observations by way of the light and fluorescence microscope revealed the characteristics of keratin variants and precursors in some necrobiotic cells which influence the peculiar properties of fibrinoid necrosis of the skin epithelia. The role of the mixed paraproteins is emphasized and reference is made to the role of a mixed paraprotein ("keratofibrinoid") which is formed in the course of the regressive process and to which the morphological changes may be attributed.


Microscopy of tonsillar smears and sections in tonsillar gonorrhoea. Microscopy of methylene-blue and Gram-stained smears from the tonsillar surface and an immunofluorescence (IF) test were carried out for 130 patients, 129 with genital and/or anal gonorrhoea, 27 of whom also had tonsillar gonorrhoea. One patient had only tonsillar gonorrhoea. 5 of the 28 patients with tonsillar gonorrhoea had acute tonsillitis and for these, agreement was found between culture, light microscopy and IF test, while only 5 of the remaining 23 patients had positive microscopy. Among the 102 patients who did not have tonsillar gonorrhoea a few false-positive light microscopies and IF tests were found. Microscopy of haematoxylin-eosin stained sections of 8 tonsils from 4 patients with recurrent tonsillitis and tonsillar gonorrhoea showed subacute tonsillitis. Methylene-blue and Gram-stained sections revealed gram-negative diplococci morphologically similar to gonococci. The bacteria were located in the superficial layers of the mucous membrane (frequently intracellularly in leukocytes) and occasionally in cellular debris in the crypts.


Five new Escherichia coli K antigens, K95, K96, K97, K98 and K100. Five Escherichia coli strains were established as antigenic test strains for five new polysaccharide K antigens: K95, K96, K97, K98 and K100. K95 to K98 served already as test strains of O antigens O75, O77, O81 and O107 respectively. F147, which is test strain of K100, had O antigen O75.


The impact of HLA-A, B or HLA-D identity on the MIF production in MLC. The relative influence on MIF release in MLC of different parts of the HLA region was studied in 1) MLCs between cells from HLA-A and B identical (probably HLA-D unidentical) unrelated persons and 2) MLCs between cells from HLA-D identical, HLA-A and B unidentical unrelated persons. It was found that the HLA-D part of the region has the major effect on the MIF production. Studies of MIF activity in supernatants from mitomycin treated cultures lent some evidence to the hypothesis that there is an extreme linkage disequilibrium between the HLA-D alleles and hypothetical separate MIF alleles. Examination of families with crossover between the HLA-A, B and HLA-D part of the region is required for confirmation of these findings.


Study of substances related to carcinoembryonic antigens, CEA-NCA and association with alpha1-antichymotrypsin. The alpha-protein which we previously detected in PCA extracts of malignant tumours, normal tissues, sera and other body fluids has been further tested. The alpha-protein was identified by immunological methods as alpha1-antichymotrypsin. Experiments by crossed immunoelectrophoresis showed that this protein cross-reacts with the betaE-protein found in the PCA extracts. The betaE-protein shows heterogeneity in molecular size, electrophoretic mobility and it has been demonstrated that it shares antigenic determinants with the CEA or betaI-molecule. Cross-reaction between betaE and other glycoproteins found in the PCA extract of normal or malignant tissue or serum was not detected.


Further characterization of protein A reactive and non-reactive subfragments of Fc from human IgG. Tryptic digests of acid-treated Fc from normal human IgG were separated into four peaks (I-IV) by gel filtration on Sephadex G-100. The second peak was further divided into two fractions (II and II'). Peak I was indistinguishable from intact Fc on electrophoresis, immunodiffusion, and reactivity to protein A. The protein A reactive fragments of fractions II, II', and III were shown to contain antigenic determinants of both the CH2 and CH3 domains, to interact with the anti-Gm (1) specific rheumatoid factor, and to fix complement. These results, together with SDS-electrophoresis, showed that protein A reactive fragments are all composed of an intact Fc chain with shorter chains covalently linked to it. The protein A non-reactive fragments of fractions II' and III were homogeneous, fixed complement and showed no interaction with the Gm (1) rheumatoid factor. These results, in addition to the observed antigenic determinants, localized the fragments to the CH2 region.


Antigenic properties of a DNA-preparation from calf thymus used for the demonstration of anti-DNA. It was attempted to evaluate passive haemagglutination of antigen coated, tanned erythrocytes as a test by which to demonstrate anti-DNA in systemic lupus erythematosus. The antigens was prepared using a minimum of procedures in order to produce a native preparation. The resulting material had most of the criteria applying to native DNA, but the protein content was about 9%. It contained a thymocyte specific component, but no demonstrable trace of bovine species antigen. The reactions between the antigen and an anti-DNA serum from a patient with suspected SLE were inhibited by DNA and DNA-histone, but not appreciably by ENA, RNA or desoxyribonucleosides. Passive haemagglutination reactions against the antigen were positively correlated to a homogeneous immunofluorescence nuclear pattern and negatively correlated to a speckled pattern. Passive haemagglutination titres against ENA and DNA antigen were not correlated. Seventy-three per cent of randomly selected sera gave either purely DNase sensitive reactions (19%) or reactions of combined sensitivity to DNase and other enzymes. Twenty-eight out of 53 sera reacting in the passive haemagglutination test reacted also in the immunofluorescence test against Chrithidia luciliae kinetoplasts. The latter reactions were DNase sensitive. It applies to both tests that DNase sensitive, but RNase resistant, reactions were well correlated, irrespective of their sensitivity to trypsin while DNase resistant or DNase and RNase sensitive reactions were not correlated. The passive haemagglutination test using a native but relatively crude DNA-preparation coated on tanned sheep erythrocytes supplemented by specificity tests with DNase and RNase treated antigen gives about the same information as the indirect immunofluorescence test against Chrithidia luciliae kinetoplasts. Furthermore, the results show that patients' sera reacting with a homogeneous nuclear pattern in the indirect immunofluorescence test may contain not only anti-DNA and anti-nucleohistone antibodies, but also antibodies to a number of non-histone chromatin associated proteins some of which contain RNA.


Encephalitogenic activity of the N-terminal part of bovine myelin basic protein in rabbits. The disease-inducing activity of the N-terminal part (residues 1-42) of bovine myelin basic protein in rabbits is localized to two regions. One encephalitogenic determined probably incorporates at least one of the lysine residues (Nos. 4 and 11); the other is within residues 20-36. It is concluded that region 1-42 shows a comparably strong degree of encephalitogenicity for rabbits.


Inhibition of the mitogenicity of the carrier molecule results in loss of immunogenicity of a hapten-LPS conjugate. Colistin methanesulfonate, a basic polypeptide similar to polymyxin E, has been shown to suppress the mitogenicity of lipopolysaccharide (LPS) from E. coli. It also inhibits the immunogenicity of a hapten-LPS conjugate. The inhibition was neither due to interference with the expression of hapten determinants, nor was it due to crossreactivity between the hapten and colistin methanesulfonate. As mitogenicity and immunogenicity was similarly affected, we conclude that activation of bursa-derived lymphocytes, in specific thymus-independent immune responses, does not take place in the absence of a mitogenic (non-Ig mediated) signal, thus supporting the hypothesis of the "one nonspecific signal" for B cell triggering.


Intracellular localization of carbonic anhydrase in the frog nephron. The intracellular distribution of carbonic anhydrase has been studied in the frog nephron by a histochemical method. The study of the detailed intracellular localization was possible by sectioning tissue embedded in plastic (Sorwall, JB-4), before staining essentially according to Hansson. This procedure preserves the intracellular structure well and the resolution is high. For light microscopy sections from 1 to 10 mum were used. For electron microscopy 0.25 mum thick sections were stained and examined in the electron microscope using an accelerating voltage of 100 kV. The highest concentrations of the enzyme seem to be localized to the cell membranes or their immediate neighbourhood. A faint cytoplasmic staining may sometimes be observed. In distal tubule cells the apical part of the cell membrane was heavily stained. Weaker staining was found at the lateral membranes and their infoldings and to some extent the basal parts of the cell membrane. In the canaliculi cells only the lateral and basal parts of the membranes were stained. This latter localization is similar to that in the parietal cells of the mammalian stomach.


Radioimmunoassays for prostaglandin metabolites. Radioimmunoassays were developed for the main plasma and urinary metabolites of PGF2alpha in the human, viz., 15-keto-13,14-dihydro-PGF2alpha and 5alpha, 7alpha-dihydroxy-11-ketotetranorprosta-1,16-dioic acid. The methods allowed assay of unextracted samples and had a sensitivity of 5 pg and 2 pg, respectively. The assays were evaluated by comparison with mass spectrometric data. Normal levels of these metabolites were estimated in a number of subjects. Levels in plasma and urine of the metabolites were followed after injection or during infusion of PGF2alpha and during administration of various antiinflammatory drugs.


Studies on the characteristics of poliovirus type 3. II. Characteristics of "hot" clones. Markers d, IST, EA1(OH)3, rct (at sub- and supraoptimal temperatures) and neurovirulence were determined for clones isolated from two lots (S2 and S3) of vaccines containing poliovirus strain Leon 12a1b. Changes of markers rct, d and neurovirulence were observed in several clones isolated from S2 vaccine. No changes were observed in IST and EA1(OH)3 markers.


Electron microscopy of surface structures of Rickettsia prowazeki stained with ruthenium red. In studying surface structures of Rickettsia prowazeki (E and Breinl strains) by ruthenium red staining, a microcapsular layer 125-165 A thick, composed of subunits 85--100 A in diameter with a periodicity of 100--120 A as well as the inner layer of the cell wall 40--60 A thick were clearly revealed. In tangential sections of cells, subunits of the microcapsular layer were found in parallel striation arrays. These structures presumably contain acid mucopolysaccharides detectable by ruthenium red staining. Besides, hitherto unreported intracytoplasmic membrane structures were detected in ruthenium red-stained rickettsiae.


Histogenesis of splenic lesions in Hodgkin's disease. Histochemical markers were used to identify the various cellular and structural components of the human spleen, and to investigate the histogenesis of the splenic lesions of Hodgkin's disease. The early lesions appear in areas near the central artery (periarterial lymphatic sheath) in the white pulp. The white pulp becomes hypertrophic. The lesions enlarge, extend into the red pulp, and compress the sinuses and the cords of Billroth. The derivations of various "histiocytes" contained with the lesions are differentiated by using cytochemical stains for lysosomal enzymes and for granulocytes. The epithelioid cells in the granulomas are rich in those lysosomal enzymes typically seen in phagocytic histiocytes, suggesting that they are indeed true histiocytes. The malignant "histiocytes," including the mononuclear Hodgkin cells, the binucleated Sternberg-Reed cells, and the multinucleated giant cells, do not contain significant amounts of lysosomal enzymes and more closely resemble stimulated lymphocytes. The splenic lesions in Hodkin's disease may be the result of a lymphocytic and histiocytic cellular response to an unknown agent, which reaches the spleen through the central artery in the white pulp.


Use of retrograde cholangiography in guiding radiotherapy of obstructive jaundice due to tumor. A patient with oat cell carcinoma of the lung which was responsive to radiotherapy developed jaundice. Endoscopic retrograde pancreatocholangiography (ERCP) revealed compression of the distal common bile duct. Radiotherapy to this area produced prompt and prolonged relief of the obstructive jaundice. The use of ERCP which led to significant palliation in a patient with known inoperable cancer makes this case worthy of note.


Identification of cells in culture.

Identification of cells in culture. Most laboratories using cells cultured in vitro maintain multiple cell lines. Such lines should be monitored for species and intraspecies characteristics to prevent invalidation of research work due to incidents of cell line cross-contamination. This report describes the results obtained when 246 cell cultures were examined for evidence of cross-contamination or mislabeling. Using species-specific antigens, isoenzyme electrophoresis, and chromosomes as markers of identity, 14% of the cultures submitted were found to be contaminated by cells of another species. Of human cell lines submitted 25% were of HeLa cell origin, as determined by 2 intraspecies markers, glucose-6-phosphate dehydrogenase and chromosome analyses. The fact that, overall, nearly 30% of the cell lines examined were incorrectly designated makes the importance of cell line monitoring self-evident.


Toxemia of pregnancy: assessment of fetal distress by urinary estriol and circulating human placental lactogen and alpha-fetoprotein levels. The efficacy of three biochemical methods for the detection of fetal distress was assessed in a prospective study of 224 singleton pregnancies complicated by toxemia. Fetal distress was diagnosed in 65 cases (29 per cent). Abnormally low urinary estriol (E3) excretion pointed out 63 per cent, low serum levels of human placental lactogen (HPL) 27 per cent, and elevated maternal serum alpha fetoprotein (AFP) 10 per cent of distressed fetuses. The efficacy of each test increased with the severity of maternal disease. The frequencies of false pathologic levels were: E3 19 per cent, HPL 0 per cent, and AFP 1 per cent of the cases with a normal fetal outcome. Although E3 was by far the most effective marker, abnormal levels of HPL and AFP provided supportive evidence for fetal distress by pointing out those cases in which E3 reading was not a false positive.


A light microscopic, autoradiographic study of axoplasmic transport in the optic nerve head during ocular hypotony, increased intraocular pressure, and papilledema. Cyclocryotherapy of the monkey eye reliably produces transient increased intraocular pressure followed by prolonged hypotony, during which papilledema occurs. Axoplasmic transport was studied while the intraocular pressure was both increased and decreased after cyclocryotherapy by autoradiography following the intravitreal injection of tritiated leucine and proline. Surgical fistulization of the anterior chamber was also used to produce hypotony and papilledema. Significant alterations of both the rapid and the slow components of axoplasmic transport were demonstrated in the nerve head during increased intraocular pressure, and in ocular hypotony with papilledema.


Pharmacologic modulation of inflammatory mediator release by rat mast cells. Current knowledge of the mechanism of inflammatory mediator release from mast cells is reviewed with particular reference to the role of cyclic nucleotides and calcium and their interrelationship with one another as defined by studies in highly purified rat peritoneal mast cells. Data are presented indicating an important role for intracellular cAMP and calcium in the mediation or modulation of release, as well as evidence for a close relationship between these two regulatory systems. Releasing agents which clearly act at the level of the plasma membrane (concanavalin A and anti-IgE antibody) are shown to differ from releasing agents that may not (48/80 and the ionophore A23187) in regard to the early cellular cAMP response, dependency of the release reaction on phosphatidyl serine, and kinetics of release. Pharmacologic modulators of release are discussed; these include: PGE1 and theophylline, which raise cAMP and inhibit release; and diazoxide, adenine, and carbachol which lower cAMP and potentiate release. Adenosine was also found to enhance release with marked effects at concentrations in the low nanomolar range.


Interaction of genes controlling two allotypes in chickens. This paper presents the results of the investigations of the newly detected antigen of chicken blood serum, called K2. It was established that the K2 antigen which was identified with isoimmune serum was a beta-globulin with the molecular weight over 200 000. The results of the genetic analysis based on sire-dam-offspring combinations seemed to indicate that the antigen under examination was controlled by a gene hypostatic to the gene controlling the previously described K1 allotype.


Removal of blood group determinants from bovine erythrocyte membranes. 3. Action of proteolytic enzymes on intact cells. Bovine erythrocyte treatment with chymotrypsin, trypsin, pronase, papain, or ficin eliminated or weakened the reactivity of 18 of the 47 blood group factors which were examined. Thirteen of the affected factors were from the B system, and one each was from the C, FV, L, M, and R'S' systems. Variation attributable to pheno-group (allele) or genotype influences was observed in the effects upon six of the factors. Ficin-treated V/V, but not F/V or F/F, cells were rapidly lysed by normal rabbit serum (complement control). Absorptions with pronase-treated V positive cells indicated that essentially all V antigenicity was removed. However, immunizations with pronase-treated V positive cells elicited V antibody production in one of two recipient cows. The numbers of antigens removed by different enzymes did not appear to be closely related to the amount of protein removed.


Alleviation of primary fibrinolysis after hypophysectomy. A case of primary hyperfibrinolysis secondary to disseminated prostatic carcinoma is presented. Transsphenoidal hypophysectomy resulted in a significant palliative response and reversal of the bleeding tendency.


Serotherapy of avian reticuloendotheliosis virus-induced tumors. Immune serum, obtained from animals that had survived sublethal challenge with RE virus, was very effective in achieving tumor cures in chickens infected with this virus. The therapeutic effect could also be obtained by the immunoglobulin fraction of immune serum. The serum did not influence the development of an unrelated malignancy. Serotherapy studies in Bx and Tx recipients indicate that the B-cell or T-cell system of the host does not significantly contribute to the curative activity of immune serum. Absorption studies show that the curative effect is mediated by antibodies to tumor-associated transplantation antigens on tumor cells, not by antiviral antibodies.


Background and practical aspects of hepatitis B surface antigen carrier state. The prevalence of hepatitis B surface antigen (HBSAg) carriers in the Europe and North America 0.1 - 0.2%, but can exceed a few per cent in South-East Asia and Africa. The HBSAg carrier state is considered to be connected with sociaohygienic conditions but it is evident that genetic factors also contribute the chronic carrier state. Practical measures concerning the treatment and management of HBSAg carriers are discussed.


Histological characteristics of chronic hepatides and primary biliary cirrhosis with special reference to orcein positive hepatocellular accumulations. The histological criteria of chronic hepatides (chronic persistent and aggressive hepatitis) and primary biliary cirrhosis are well characterized and documented in the literature. Histology forms the diagnostic basis for chronic hepatides. Diagnostic changes are seen, however, only in some cases of primary biliary cirrhosis (PBC) and only in the early stages of the disease. Difficulties are met especially in differentiating PBC from chronic aggressive hepatitis (CAH). There is a considerable histological overlap and mixed forms occur. Orcein-positive material, which is a copper-binding protein with high content of sulphydryl groups, accumulates in liver cells in long standing cholestatic liver diseases and can be demonstrated histochemically in routine biopsy specimens. It is seen in PBC in at least 70% of biopsy specimens which confirms the biliary nature of the disease. In CAH orcein positive material can be demonstrated in 20% of specimens but only from patients who also have features of PBC. This group of patients may therefore have both biliary disease are hepatocellular damage, and can be separated from CAH by the orcein method.


Clinical correlations and significance of orceinpositivity in chronic active hepatitis and primary biliary cirrhosis. Clinical, biochemical and immunological variables were analyzed in 30 patients with orcein-negative (ON) chronic active hepatitis (CAH), 4 patients with ON primary biliary cirrhosis (PBC), 8 patients with orcein-positive (OP; intracellular copper-binding protein seen histologically in liver biopsy specimens) CAH and 15 patients with OP-PBC. A marked elevation of serum bile acids, alkaline phosphatase, leusine aminopeptidase, gammaglutamyl transpeptidase and cholesterol concentrations, and highly pathological BSP Tm values were characteristic for OP-cases. In addition the faecal fat level was increased and bile acids decreased in OP-cases. Serum levels of IgG or IgM and the occurrence of smooth muscle, mitochondrial or glomerular antibodies were identical in ON- and OP-CAH as well as also in ON- and OP-PBC. 49 patients were treated with a combination of prednisone and azathioprine from 4 to 72 months (mean 22). 26 patients with ON-CAH responded biochemically and morphologically to the treatment. No treatment failures were found in ON-CAH. In contrast treatment failure was confirmed in every treated OP-PBC or OP-CAH. The results suggest that orcein-positivity indicates a poor response to prednisone-azathioprine treatment of CAH.


The prevention of viral hepatitis.

The prevention of viral hepatitis. Studies with heat-inactivated MS-2 serum have indicated that active immunization against hepatitis B infection can be achieved even though the virus has not been cultivated in tissue culture. The development of new biophysical, biochemical and immunological techniques has increased knowledge of the hepatitis B virus and its associated antigens. In the wake of these recent developments the accumulating evidence indicates that active immunization could be achieved by the use of purified hepatitis B surface antigen or by one of its polypeptides or glycolipids.


Method for reproducible large-volume production and purification of Rauscher murine leukemia virus. Rauscher murine leukemia virus was produced in roller-bottle cultures of chronically infected JLS-V9 cells. Virus from this culture fluid was concentrated and purified by two semi-isopycnic bandings in sucrose gradients. Virus material obtained from young, nonconfluent cultures (early-harvest virus) yielded products characteristically containing endogenous ribonucleic acid-dependent deoxyribonucleic acid polymerase with high specific activity (400 to 1,000 pmol of [3H]thymidine 5'-triphosphate incorporated per milligram of protein per hour). Fluids obtained from older confluent cultures (late-harvest virus) yielded products with endogenous ribonucleic acid-dependent deoxyribonucleic acid polymerase with little or no specific activity (200 pmol or less of [3H]thymidine 5'-triphosphate incorporated per milligram of protein per hour), but with higher virus particle counts and greater amounts of protein and gs antigen than the early-harvest products.


Penetration and comparative metabolism of leptophos in susceptible and resistant houseflies. The metabolism and rate of penetration of leptophos (O-methyl O-4-bromo-2,5-dichlorophenyl phenylphosphonothioate) was determined in a susceptible strain and a strain of housefies which was 50-fold resistant to leptophos. Penetration of leptophos into resistant flies was substantially slower than into susceptible flies but large differences in metabolism, both quantitatively and qualitatively, were not observed. No difference was observed in the sensitivity of flyhead and thorax acetylcholinesterase to leptophos-oxon in vitro, and tolerance to leptophos by the resistant strain is explained in terms of decreased rates or penetration and minor differences in metabolism.


Insecticide movement following application to crevices in rooms. The movement of chlorpyrifos and diazinon from cracks and crevices in empty dormitory rooms with and without forced air movement to houseflies and plates in non-target areas was studied. Aluminum pie plates in rooms with the high concentrations of an insecticide, no forced air movement, and at the earlier sampling interals contained more insecticide. Insecticides were not detectable at 48 hr. Increased housefly movement occurred simultaneously with the initiation of insecticide application in rooms with forced air and 1 min later in rooms with no forced air. Mortality of houseflies exposed during and at 5 hr after insecticide application was greater for males than females in all tests except one; at the high insecticide concentrations; and at successive time intervals through the 24-hr interval after their transfer from the treated rooms to holding rooms. Houseflies in cages near the ceiling, exposed to an insecticide, and forced air movement, exhibited greater mortality than those in cages on the floor, while the reverse was true for houseflies in rooms with no forced air movement.


The evolution of the concept of the use of surgical castration in the palliation of breast cancer in pre-menopausal females. The understanding of the association of mammalian ovarian function with lactation was common knowledge to dairy-men early in the 19th century or earlier. This thesis reviews the application of these empiric observations to lactation in humans by Beatson of Glasgow, supported by his preliminary laboratory investigations. Beatson noted the cellular anatomic similarity between pre-gestational breast and carcinoma. This study reviews the progressive development of the successful clinical application of castration for the control of inoperable breast cancer by Beatson in pre-menopausal women in 1895. The review terminates with its leading to the monumental work of Huggins in 1941.


Palliation of infants with truncus arteriosus through creation of a pulmonary artery ostial stenosis. A surgical procedure for palliation of infants with truncus arteriosus and excessive pulmonary blood flow (types I, II, and III) is described. The technique involves the production of ostial stenosis of the pulmonary artery from within the truncus using cardiopulmonary bypass. This method has advantages over pulmonary artery banding, which in general has produced disappointing results.


[Effect of long-lasting noise on the plasma concentration of catecholamines, glucocorticosteroids and PBI in pigs]. Exposure of pigs with a body mass of 25-30 kg to long-lasting motor noise (108 dB, 60-8000 Hz, 72 hours) results in a considerable increase of catecholamine concentration in the plasma. Coincidently there is a sharp continuous decrease of the 11-OHKS in the plasma during the whole period of exposure to noise. Immediately after stopping the acoustic irritation the concentration of plasma corticosteroids increases again. The electroencephalogram (EEG) giving evidence of inhibition signs during exposure to noise an adaptation effect of the information processing system is concluded. The increase of the PBJ during the period of acoustic stress points to an increased function of the thyroid gland but simultaneously it is considered to be an expression of the elevated adrenergic situation. By reason of the strong deviation of the three observed regulatory systems must be concluded that longer-lasting exposure to noise with the applied quality and intensity represents a stress situation for pigs having noise character as established for men. A possible influence on the productivity of the pork type pig by permanent acoustic stress is indicated.


Conditions favoring immunological enhancement of a murine allogeneic lymphoma. An in vivo model was used which permits the growth of AKR lymphoma allografts inoculated within a glass cylinder subcutaneously implanted in BALB mice. Pretreatment of the host with acellular tumor extracts or tumor cells enclosed within a diffusion chamber significantly increased tumor incidence. On the contrary, donor spleen extracts did not alter tumor incidence while viable spleen cells within a diffusion chamber even prevented tumor development. It can be concluded that in this model a condition of maximal tumor enhancement can be attained with soluble tumor antigen but not with normal spleen extracts.


Primary immune response to sheep red blood cells in mice treated with immunoregulatory alpha-globulins (IRA). Alpha-Globulins from human and bovine sera or from mouse ascitic fluid were separated by ion-exchange chromatography on DEAE-cellulose into fractions A, B, and C. Fractions A and B had no immunosuppressive activity; fraction C injected into mice at the time of antigen administration, but not later, significantly reduced the number of anti-SRBC plaque-forming cells in the spleens of experimental animals. Single high dose inhibited the response better than the same doses given on 4 consecutive days. It is concluded that a critical level of alpha-globulins is necessary to render lymphocytes hyporesponsive, and when once stimulated by antigen they become less susceptible to alpha-globulin action.


Anti B and T cell activity of antiserum to spleen cells of athymic (nu/nu) mice: evidence for surface antigenic differences between functionally distinct subclasses of T and B lymphocytes. A rabbit antiserum obtained by immunization with spleen cells of nu/nu mice, known to be deficient in thymus and T cells but not in T cell precursor, possesses two different specificities. In addition to activity against bone marrow-derived lymphocytes (B cells), as revealed by inhibition of antibody forming cells and myeloma cells, the antiserum (Ra-nu/nu) has strong activity against cytotoxic T cells but not against helper T cells. Differences in sensitivity to antiserum treatment between anatomically and functionally distinct B cells point to surface antigenic differences among subclasses of B cells.


Comparative study of the electrophoretic mobility of the RNA of influenza parent and recombinant strains. The electrophoretic mobility of the RNA of Influenza viruses A/WSN, A/Singapore and their antigenic recombinants X-7 and X-9 was investigated. The genome of each virus studied consisted of seven pieces of RNA. The electrophoretic profile of the influenza virus A/WSN RNA differed from that of A/Singapore but resumbled that of the recombinant X-9 genome. The essential differences were connected with the properties of the fifth fragment of the RNA. The molecular weight of this RNA species of influenza virus A/WSN and X-9 was 5.4 X 10(5) AND 5.3 X 10(5) daltons (d) respectively. The molecular weight of the corresponding component of the influenza viruses A/Sinapore and X-7 RNA was 6.2 X 10(5) and 6.3 X 10(5)d respectively.


Effect of Ca++ on the stability of influenza virus neuraminidase. The neuraminidases of different strains of influenza virus varied in their stability at 37 degrees C. The enzymes of the strains with N1 neuraminidases were found to be unstable during incubation at 37 degrees C whereas the enzymes of the strains with the N2 neuraminidases were stable. Among the strains with N2 neuraminidases, the enzymes of some strains were inactivated during dialysis at 37 degrees C whereas the enzymes of others were stable. This observed loss of enzyme activity during dialysis at 37 degrees C was not restricted to a single substrate as the same loss of enzyme activity was observed irrespective of the size of the substrate used in the assay. The enzymically inactive neuraminidase was found to be non-antigenic and non-immunogenic. The inactivation of the enzyme could be prevented by the addition of Ca++ but not Mg++. Out results suggest that Ca++ is essential for the stability of the enzyme at 37 degrees C. The results would also suggest that the enzymic, antigenic and immunogenic sites are either the same or very closely situated on the surface of the neuraminidase molecule.


A continuous-wear hydrophilic lens. Prophylactic topical antibiotics. Seventeen patients, who were fitted with a continuous-wear hydrophilic lens for 12 weeks, received topical neomycin sulfate-polymyxin B sulfate-gramicidin (Neosporin) eyedrops in one eye only at a frequency of 3.7 times per day. The continuous wear of a hydrophilic lens in and of itself did not after normal bacterial flora, and the use of the antibiotic drops did not alter normal conjunctival flora. In addition, the eyedrops were not more effective when used in conjunction with the hydrophilic lens and did not appear to produce resistant organisms during the 12-week study. In normal eyes, a continuously worn hydrophilic soft contact lens that is well tolerated does not adversely affect the bacterial flora and the addition of neomycin sulfate-polymixin B-gramicidin drops at a frequency of 3.7 times per day does not appear to be beneficial or necessary.


Density of mucous glands in the normal adult nasal septum. The entire mucosa from 13 normal nasal septa was stained by the PAS-alcian blue whole-mount method, and the mean density of glandular orifices in 13 localities was determined quantitatively by counts in 4 mm2 fields. The median density within the entire material was 8.4 glands/mm2, with an interindividual variation of +/- 1 gland/mm2. The median number was 15,900 glands, range 10,800-21,800. The distribution of the glands is regular, with little variation between the localities. Density is somewhat, but significantly lower in the posterior quarter of the septum, and there was a significant increase in density into the infero-superior direction.


[Meatoplasty with allogenic preserved cartilage. Histomorphological model in animal experiments (author's transl)]. The reconstruction of the posterior wall of the external acoustic meatus destroyed by surgery or a cholesteatoma is necessary for the complete restoration of function. For this purpose, allogen cartilage preserved in thiomersalate has been transplanted on the anterior wall of the air-containing mastoid bone in 160 clinical cases. The histomorphological reaction of the recipient to unilaterally uncovered transplant material has previously been unknown. In 10 guinea pigs, 15 fenestrated bullae (hypotympanum equivalents) were closed with cartilage transplants fixed in place with tissue adhesive. After a period varying between 7 1/2 and 11 months, the walls of the bullae with the transplant were then examined histomorphologically. Despite the variations in the period in situ, the avital cartilage transplant, which remained largely constant in volume and shape, was always to be found. It was tolerated without reaction and was "enveloped"--also on the primarily uncovered surface--by a capsule of bone and connective tissue. In no case was rejection observed. Marked foreign-body reaction was, however, found in the vicinity of the tissue adhesive, which was absorbed only slowly. The clinically well-known tissue-compatible and complication free behaviour of the allogenic cartilage transplant preserved in thiomersalate, is thus explained by the histomorphological findings. In contrast, the fact that marked foreign-body reaction was demonstrable even after almost one year, suggests that tissue adhesive should not be used.


The conquest of smallpox.

The conquest of smallpox. Smallpox has been a dreaded disease for thousands of years. Recurrent epidemics traversed the known world leaving a trail of death, disfigurement, and disability unparalleled by the other great plagues. Fear of smallpox provoked the first important attempts to prevent epidemic disease by a primitive form of immunization called variolation. In 1796, Edward Jenner developed the first effective vaccine against an infectious disease by using cowpox virus to prevent subsequent infection with smallpox. The discovery of this vaccine made possible the conquest of smallpox, a task that will probably be completed this year.


Quantitation of casein messenger ribonucleic acid sequences using a specific complementary DNA hybridization probe. Two highly purified rat casein mRNA fractions were used as templates to synthesize complementary DNA (cDNA) hybridization probes using RNA-directed DNA polymerase isolated from avian myeloblastosis virus. Both of the probes selectively hybridized to RNA isolated from lactating mammary tissue, but not to poly(adenylic acid)-containing rat liver RNA. An analysis of the kinetics of hybridization of the cDNA derived from the 15S casein mRNA (cDNA12S) with their individual mRNA templates indicated that greater than 90% hybridization occurred over a R0t range of one and one-half logs with R0t 1/2 values of 0.0023 and 0.0032 mol s l.-1, respectively. Compared with the total RNA isolated from lactating mammary tissue, these values represented a 166- and 245-fold purification, respectively, of these individual mRNA fractions. Using the 15S casein mRNA as a template, two probes of different lengths and specific activities were synthesized. The deoxyribonucleotide and mRNA concentrations and the temperature of incubation were optimized to obtain either a high specific activity cDNA probe, 330 nucleotides long, which represented approximately 25% of the mRNA or a lower specific activity preparation containing some complete cDNA copies, 1300 nucleotides in length. The Tm of the longer cDNA15S-15S mRNA hybrid was 88.5 degrees C, while that of the short cDNA15S-RNA hybrid was 82.5 degrees C. Following this initial characterization, the cDNA15S probe was utilized for three separate determinations: (1) Analysis of the sequence divergence between mouse and rat casein mRNAs. It was observed that the rate of hybridization of heterologous rat cDNA15S-mouse casein mRNA was only 20% that of the homologous rat cDNA15S-rat casein mRNA hybridization. The resulting heterologous hybrid displayed approximately 17% mismatching compared with the homologous hybrid. (2) Determination of the gene dosage for casein mRNA in normal and malignant mammary cells. In this study, an analysis of the kinetics of hybridization of the high specific activity cDNA15S probe with an excess of DNA isolated from lactating mammary tissue, carcinogen-induced mammary tumors, or rat liver indicated that casein mRNA was transcribed from the nonlification or deletion was observed during tumor formation or the process of mammary differentiation. (3) Quantitation of casein mRNA sequences during normal mammary gland development. RNA excess hybridizations were performed using RNA extracted from either pregnant, lactating, or regressed rat mammary tissue. The concentration of casein mRNA molecules/alveolar cell was found to increase 12-fold from 5 days of pregnancy until 8 days of lactation and then declined to approximately 2% of the maximal level of 79 000 molecules/cell by 7 days after weaning. A coordinate increase was observed in casein mRNA sequences detected by cDNA hybridization and mRNA activity measured in a cell-free translation assay.


Kinetic studies of drug-dinucleotide complexes. Three classes of kinetic behavior are observed in the complexes of actinomycin or ethidium with deoxydinucleotides. First, the initial dinucleotide binding to form a 1:1 complex is a rapid bimolecular process, whose rate could be measured for combination of actinomycin with d(pTpG) d(pGpT), d(pGpA), d(pGpG) d(pCpGpG), and d(pCpG) andfor combination of ethidium with d(pGpC). Second, with one exception, all reactions in which a second dinucleotide is added to form a 2:1 dinucleotide-drug complex are limited by a first-order step at high concentration. This class includes the combination of actinomycin with all dinucleotides tested except d(pGpC), and the reaction of ethidium with nucleotides of complementary sequence pyrimidine-purine, such as d(pCpG). The final class is the special case of d(pGpC) interacting to form a 2:1 complex with actinomycin. Third-order kinetics is observed, with no evidence for a first-order, rate-limiting step.


Anthroyl stearate as a fluorescent probe of chloroplast membranes. 1. A reversible light-induced enhancement of the fluorescence of a "hydrophobic fluorophore", 12-(9-anthroyl)-stearic acid (anthroyl stearate), is observed with chloroplasts supporting phenazine methosulfate, cyclic or 1,1'-ethylene-2,2'-dipyridylium dibromide (Diquat) pseudo-cyclic electron flow; no fluorescence change is observed when methyl viologen or ferricyanide are used as electron acceptors. The stearic acid moiety of anthroyl stearate is important for its localization and fluorescence response in the thylakoid membrane, since structural analogs of anthroyl stearate lacking this group do not show the same response. 2. This effect is decreased under phosphorylating conditions (presence of ADP, Pi, Mg2+), and completely inhibited by the uncoupler of phosphorylation NH4Cl(5-10mM), as well as the ionophores nigericin and gramicidin-D (both at 5 - 10(-8)M). The MgCl2 concentration dependence of the anthroyl stearate enhancement effect is identical to that previously observed for cyclic photophosphorylation, as well as for the formation of a "high energy intermediate". The anthroyl stearate fluorescence enhancement is inhibited by increasing concentrations of ionophores in parallel with the decrease in ATP synthesis, but is essentially unaffected by specific inhibitors (Dio-9 and phlorizin) of photophosphorylation; thus, it appears that anthroyl stearate monitors a component of the "high energy state" of the thylakoid membrane rather than a terminal phosphorylation step. 3. The light-induced anthroyl stearate fluorescence enhancement is suggested to monitor a proton gradient in the energized chloroplast because (a) similar enhancement can be produced by sudden injection of hydrogen ions in a solution of anthroyl stearate; (b) when the proton gradient is dissipated by gramicidin or nigericin light-induced anthroyl stearate fllorescence is eliminated; (c) when the proton gradient is dissipated by tetraphenylboron, light-induced anthroyl stearate fluorescence decreases, and (d) light-induced anthroyl stearate fluorescence change as a function of pH is qualitatively similar to that observed with other probes for a proton gradient (e.g. 9-aminoacridine). Furthermore, anthroyl stearate does not monitor H+ uptake per se because (a) the pH dependence of H+ transport is different from that of the anthroyl stearate fluorescence change, and (b) tetraphenylboron, which does not inhibit H+ uptake, reduces anthroyl stearate fluorescence. Thus, anthroyl stearate appears to be a useful probe of a proton gradient supported by phenazine methosulfate of Diquat catalyzed electron flow and is the first "non-amine" fluorescence probe utilized for this purpose in chloroplasts.


Circular dichroism of intermediate subviral particles of reovirus. Elucidation of the mechanism underlying the specific monovalent cation effects on uncoating. 1. Circular dichroic (CD) spectra of purified intermediate subviral particles of reovirus were determined in the presence of different monovalent cations. 2. The CD spectra reveal that reo intermediate subviral particles can exist in two conformationally different forms. The two forms are readily distinguished by comparison of their ellipticities in the wavelength regions 210 nm and 220 nm, with a Na+-induced form exhibiting a reduced negative ellipticity relative to a Cs+-induced form. 3. The transition between the Na+- and Cs+-induced forms is reversible by manipulation of the species of monovalent cation present and appears to be temperature independent. 4. Temperature variation studies on dilute suspensions of particles indicate that the Na+-induced form is stable, whereas the Cs+-induced from undergoes a second transition, temperature dependent and irreversible, to become a viral core. 5. A model is presented relating these observations to the known properties of reovirus uncoating and transcriptase activation.


Purification and characterization of gibbon ape leukemia virus DNA polymerase. An RNA directed DNA polymerase was purified over 2500 fold from gibbon ape leukemia virus by successive column chromatography on Sephadex G100, DEAE cellulose, phosphocellulose and hydroxyapatite. The purified DNA polymerase has a molecular weight of 68 000, a pH optimum of 7.5, a Mn2+ optimum of 0.8 mM, and KCl optimum of 80 mM. The purified enzyme transcribes heteropolymeric regions of viral 60-70 S RNA isolated from avian myeloblastosis virus, Rauscher murine leukemia virus and simian sarcoma virus and it is inhibited by antiserum prepared against either gibbon ape leukemia virus or simian sarcoma virus DNA polymerases.


Interaction of rifamycins with mammalian nucleic acid polymerizing enzymes. Procedures were established for the isolation and partial purification of DNA polymerase, RNA polymerase and poly(A) polymerase activities from the cytoplasm and nuclei of NIH-Swiss mouse embryos. Based on the elution pattern of these enzyme activities from DEAE-cellulose and phosphocellulose columns in Tris-HCl buffer, pH 8.0, the apparent basicities of the enzymes can be arranged as follows: cytoplasmic(C) poly(A) polymerase greater than (C)DNA polymerase beta greater than (C)DNA polymerase alpha and nuclear(N) poly(A) polymerase greater than (N)DNA polymerase greater than (N)RNA polymerase I greater than (N)RNA polymerase II. Twenty rifamycins, including rifamycin B, rifamycin S, rifamycin SV, and rifamycin SV derivatives, were examined for their ability to inhibit the above mentioned nucleic acid polymerizing enzymes and Simian sarcoma virus type I (SSV-1) reverse transcriptase. Rifamycin SV 3'-formyldiphenylhydrazone, rifamycin SV 3'-formyl-n-octyloxime (AF/013) and rifamycin SV 3'-formyldiphenylmethyloxime (AF/05) inhibited all the tested enzyme activities. Rifamycin SV 3'-formylpropylphenyloxime (AF/015) inhibited cellular nucleic acid polymerase activities but not SSV-1 DNA polymerase activity. Rifamycin SV 3'-formyldinitrophenylhydrazone (AF/DNFL) strongly inhibited reverse transcriptase activity but did not inhibit cellular DNA polymerase activities. AF/DNFI slightly inhibited RNA and poly(A) polymerase activities. Rifamycin SV 3'-formyldipropylhydrazone (AF/DPI) and 2,6-dimethyl-4-N-benzyldemethyl-rifampicin (AF/ABDMP) slightly inhibited reverse transcriptase activity but did not inhibit cellular nucleic acid polymerase activities. Active rifamycin derivatives inhibited enzyme reactions by interacting with the enzyme proteins. Nascent polynucleotide chain elongation continued although at a reduced rate in the presence of inhibitor. The addition of increasing concentrations of nonionic detergent (Triton X-100) to rifamycin-inhibited enzyme reactions fully restored enzyme activities. The presence of highly lipophilic 3'-side chains on active rifamycins and the reversibility of enzyme inhibition by Triton X-100 suggest that the tested nucleic acid polymerizing enzymes may have hydrophobic regions with which inhibitory rifamycins interact.


A new estimate of human ribosomal gene number. Radioactively labelled DNAs (5 X 10(6) cpm/mug) complementary to human 18 S and 28 S ribosomal RNA were synthesized using RNA-directed DNA polymerase (EC 2.7.7.7). These complementary DNAs were used to measure human ribosomal gene numbers by two independent methods, both of which indicated numbers at least four-fold lower than those previously reported. First, the kinetics of the annealing of the complementary DNAs with total human placental DNA indicated that the number of both 18-S and 28-S ribosomal genes per haploid genome is approximately 50. Second, saturation experiments in which a constant amount of DNA was annealed with increasing amounts of complementary DNA also indicated that the number of 28 S ribosomal RNA genes in human placental and spleen DNA is is about 50 per haploid genome.


Latent homology of murine lymphoid antigens revealed through the complement mediated absorption of xenogeneic antiserum. Xenoantiserum against C57BL/6 mouse spleen cells was produced by immunization of rabbits. Such antiserum displayed cytotoxic activity for several strains of mice. Standard "in vitro" absorption removed species specific antibody revealing antibody directed against the C57BL/6 strain. When the xenoantiserum was absorbed with DBA/2 spleen cells in the presence of complement, the binding of anti-C57BL/6 was increased. Such data suggest that xenoantibodies directed against spleen cells possess a broader capacity to react with cross-strain lymphoid antigens than previously described.


Antibody-like activity to polyacrylic acid in normal rabbit sera. Most unimmunized rabbit sera agglutinate acrylic particles coated with polyacrylic acid. The factor responsible for the agglutinating activity turned out to be a 2-mercaptoethanol-resistent immunoglobulin, present in low concentration. It was tentatively classified as "natural antibody". This is assumed to have arisen upon previous contact with naturally occurring cross-reacting polyanions.


Urinary excretion of total protein, albumin and beta2-microglobulin during exercise in adolescent diabetics. Urinary total protein, albumin and beta2-microglobulin excretion during exercise on a bicycle ergometer was determined in a group of 7 adolescent male diabetics and in a comparable control group. There was no difference, neither in the diabetic group nor in the control group, in the excretion of these proteins under the provocative effect of physical exercise. These results suggest exercise may not always give a positive response and is not a discriminative function in reliable studies of renal function.


Leukocyte cytochemical reactions in preterm and small for date babies. Four cytochemical reactions widely used for hematological diagnosis (Myeloperoxydase, leukocyte alkaline phosphatase, PAS and Sudan Black) were carried out in 46 infants admitted into a neonatology unit (14 preterm babies, 19 small for date newborn infants (SDB) and 13 preterm with retarded intra-uterine growth). The MPO, polymorphonuclear PAS and Sudan Black levels are comparable in the three groups of infants and the same as in normal full-term newborn babies. On the other hand, the LAP level, identical in preterms and SDB, is lower than in full-term infants but higher than in adults. Lymphocyte PAS value increases progressively with age. Better interpretation of results would be gained with knowledge of normal ranges for these four parameters in children. The authors suggest that these reactions should be used in the field of general metabolic diseases, susceptibility to infection and genetic diseases with or without chromosal abnomaly.


[Isolation and characterization of pregnancy associated alpha2 glycoprotein]. A method using immunoadsorbents for the isolation of pregnancy-associated alpha2-glycoprotein (alpha2-PAG) from the extract of human placentae is described. The physical properties and the chemical composition of the purified protein are determined: alpha2PAG sediments with 11,5 S, has a molecular weight of 360 000 daltons and is composed of subunits having a molecular weight of 180000, which are held together by disulfide bonds. The isoelectric point was found to be pH 4,7 and the extinction coefficient (E1%1cm) was determined to be 9,7 at 277 nm. The carbohydrate content of the molecule amounts to 12,1% (hexose 6,0%, hexosamine 3,7%, fucose 0,06%, sialic acid 2,4%). An analysis of the amino acids is reported, too. The purified alpha2PAG was used to determine the absolute concentrations of this protein in a reference standard and in sera.


Joint air pollution sampling program in twin cities on the U.S.-Mexico border. Ciudad Juárez (Chihuahua) and El Paso (Texas), two cities on the U.S.-Mexico border, form a single environmental system in which the same natural resources, especially air and water, are shared. It also constitutes a single metropolitan area which is characterized by high rates of pipulation growth, economic development, and urban expansion, all these factors mitigating against air quality. Early in 1972 the health authorities in El Paso and Ciudad Juárez initiated a joint air pollution sampling program with assistance from the Pan American Health Organization. The nearby city of Las Cruces (New Mexico) was later included in the program as well. Activities are carried out in accordance with a document entitled "Bases of Cooperation." The guiding criteria of the program are: functional simplicity, operational economy, and complementarity with other sampling programs conducted by the participating services. An Air Pollution Control Subcommittee is responsible for execution coordination of the program. Three studies are currently underway to determine levels of dust pollution in the air. A fourth study is aimed at measuring sulfur dioxide levels through the use of sulfation plates. The results collected reveal concentrations of particulates in the ambient air levels higher than the U.S. Federal primary standards. The program should be expanded to include the study of other pollutants and a joint inventory of emissions. In this way criteria on air quality may be established and joint plans of action and strategies drawn up for the control of air pollution in this important area.


Pharmacological properties of N-(3',4'-dimethoxycinnamoyl) anthranilic acid (N-5'), a new anti-atopic agent. 1 N-(3'-4'-dimethoxycinnamoyl) anthranilic acid (N-5') exhibited a dose-dependent, potent inhibition of the passive cutaneous anaphylaxis (PCA) mediated by homocytotropic antibodies (HTA), which was hardly affected by anti-inflammatory agents such as phenylbutazone, indomethacin and prednisolone at any dose used. The HTA-induced PCA was significantly inhibited by combined treatment with diphenydramine and cyproheptadine. 2 Doses of N-5' which potently inhibited HTA-induced PCA inhibited only slightly the heterologous PCA produced by anti-bovine serum albumin (BSA) rabbit serum. This heterologous PCA was clearly inhibited by phenylbutazone, indomethacin and prednisolone. Diphenydramine and cyproheptadine, singly or combined inhibited the heterologous PCA only slightly. 3 The increased vascular permeability caused by histamine and 5-hydroxytryptamine was significantly inhibited by diphenyldramine or cyproheptadine, but not by N-5' and the anti-inflammatory agents used. 4 N-5' 150 mg/kg orally inhibited rat paw oedema induced by carrageenin by about 26% while phenylbutazone, indomethacin and prednisolone produced significant inhibition. 5 N-5' at concentrations of 100 and 1000 muM significantly inhibited (by about 52% and 95%, respectively) the histamine release from rat peritoneal cells induced by HTA; 10 muM N-5' had little effect. Histamine release was inhibited by phenylbutazone or indomethacin at 1000 muM but not at 100 muM. Prednisolone had no effect on histamine release at any of the concentrations used. 6 These findings suggest that the inhibition of the HTA-induced PCA by N-5' may be due to inhibition of histamine release and is clearly different from the actions of anti-inflammatory agents such as phenylbutazone, indomethacin and prednisolone.


Clinical use of combined bleomycin and radiation therapy for head and neck tumours and testicular cancers. The Bleomycin advantages are resumed with special reference to the use in combination with irradiation. More vigorous effect were seen in patients given Bleomycin one hour before irradiation twice to three times weekly than after irradiation alone. For head and neck tumours the effect of two weeks Bleomycin treatment alone showed 40 per cent distinct shrinkage and in this group 73 per cent of the patients were tumour-free after a full course of radiotherapy, compared to 38 per cent in the group showing no response to Bleomycin-pretreatment. Moderately differentiated squamous cancer showed just as good response as highly differentiated tumours. The one year survival using subsequent treatment of Bleomycin and irradiation, compared to an historical material showed improved survival rate for T3 laryngeal cancers, as well as all stages of testicular carcinomas given simultaneous Bleomycin twice weekly under the full course of postoperative irradiation to regional lymph nodes.


Efferent connections of the septal area in the rat: an analysis utilizing retrograde and anterograde transport methods. Experiments were performed by either retrograde (horseradish peroxidase histochemistry) or anterograde ([3H]leucine radioautography) transport methods to determine the efferent connections of the septum in the rat. It was observed that the dorsal septum projects to the lateral preoptic area, lateral hypothalamus, periventricular hypothalamus and midline thalamus. Fibers from the ventral half of the septum project topographically to the hippocampal formation, thalamus, hypothalamus and midbrain. Specifically, neurons located along the midline in the vertical limb of the diagonal band project through the dorsal fornix to all CA fields of the dorsal hippo campus and adjacent subicular cortex. Other fibers from this region project through the stria medullaris to the medial habenular nucleus and anteromedial nuclhe pars posterior of the medial mammillary nucleus. Cells located immediately lateral to the midline in the vertical limb of the diagonal band project through the medial part of the fimbria to all CA fields of the posterior hippocampus and adjacent subicular cortex. Other fibers which originate from this region project through the stria medullaris to both the medial and lateral habenular nuclei and the paratenial nucleus of the thalamus, and through the medial forebrain bundle to the pars posterior of the medial mammillary nucleus. Cells located in the intermediolateral septum project through the lateral part of the fimbria to all CA fields of the ventral hippocampus and adjacent subicular and entorhinal cortices. These cells also send fibers through the stria medullaris to the lateral habenular nucleus and mediodorsal thalamic nucleus. Other axons arising from these cells descend through the medial forebrain bundle to terminate in a region dorsal to the interpeduncular nucleus. Fibers from the most lateral part of the ventrl septum (i.e., bed nucleus of the anterior commissure) project through the stria terminalis to the ventral subiculum. In addition, cells located in the horizontal limb of the diagonal band project massively to the pars posterior of the medial mammillary nucleus and the ventral tegmental area, and amygdala.


Banding analysis of the somatic chromosomes of the domestic dog (Canis familiaris). The chromosomes of the domestic dog (Beagle) were investigated by several different staining techniques. G-banding, Q-banding, and the bis-benzimidazol derivative Hoechst 33258, make possible the identification of all 39 chromosome pairs. Constitutive heterochromatin (C-bands) was present on a few chromosomes as distinctive, large stained areas; on the other autosomes there was little or no heterochromatin detectable.


Differential fluorescence of sister chromatids with 4'-6-diamidino-2-phenylindole. Differential fluorescence of sister chromatids and sister chromatid exchanges (SCE) in chromosomes from human lymphocytes grown two replication cycles in medium containing 5-bromodeoxyuridine can be detected by fluorescence microscopy after staining with 4'-6-diamidino-2-phenylindole (DAPI). The DAPI fluorescence appears to be more stable than that of the dye 33258 Hoechst and may provide a more sensitive method for the detection of SCE.


Persistence of transferable drug resistance in the lactose-fermenting enteric flora of swine following antimicrobial feeding. Six groups of swine (85 animals) were fed a combination of antimicrobial drugs (sulfamethazine 100 g/ton, chlortetracycline 100 g/ton and penicillin 50 g/ton). After two weeks the antimicronial drugs were removed from the diet of two groups (28 animals). These swine were compared to four groups fed the medicated diet to determine the effect of duration of treatment and degree of animal isolation on the persistence of resistance in lactose-fermenting enteric organisms. The degree of resistance to penicillin, oxytetracycline, dihydrostreptomycin and neomycin as determined by minimum inhibitory concentrations and the incidence of resistant organisms were examined during and after antibiotic feedings. Ninety-two percent or greater of all isolates tested during and after treatment had minimum inhibitory concentrations for oxytetracycline of greater than 100 mug/ml. Thirty-two weeks after cessation of dietary antibiotic, resistance to oxytetracycline and dihydrostreptomycin remained at 100% and 89% respectively. Variation in degree of contact between swine receiving medicated feed and those receiving nonmedicated feed was not sufficient to reduce the incidence of resistance to oxytetracycline or dihydrostreptomycin in all animals. Factors influencing persistence of resistant enteric organisms are discussed. Addition of the antimicrobials to the ration resulted in significantly greater weight gains for treated animals than for the controls but did not alter feed conversion.


Friedreich's ataxia: observations with Q and G banding of human chromosomes. No chromosomal anomaly was found in 15 cases of typical Friedreich's ataxia and three cases of atypical recessive ataxia studied with Q and G banding techniques. No difference between frequency of chromosomes gaps or breakages was noted amongst patients with Friedreich's ataxia and controls.


Living with the dying: use of the technique of participant observation. Through participant observation, questions concerning optimal care of dying patients and needs of their families were answered. A general surgical ward and a palliative care unit were the sites of observation. The observations support the belief that a palliative care unit, specifically designed to meet the known needs of dying patients and their families, is preferable to a general surgical ward. The main findings, of less concern in the palliative care unit than on the surgical ward, were the following: the importance of patient-to-patient support; the discomfort of sick-role behaviour; the impersonal and sometimes intimidating nature of patient care; the limitation of the patient's need (as a person) to give as well as to receive; and the value of families, student nurses and volunteers in total care. These findings emphasize the importance of personal interest in relieving the distress suffered by many terminally ill patients.


Phase II study of mitomycin-C, vincristine, and bleomycin in advanced squamous cell carcinoma of the uterine cervix. Utilizing the stathmokinetic principle of timed vincristine and bleomycin, we combined these two agents with Mitomycin-C. The dose schedule included vincristine 0.5 mg/m2 intravenously (i.v.) geginning on day 1 and repeated twice weekly for 12 weeks; each injection was followed in 6-12 hours by bleomycin 6 mg/m2 for 12 weeks. Mitomycin-C was administered as a 20 mg/m2 bolus beginning on day 2 and repeated at 6-week intervals. Thirty patients were entered into this study, 27 were fully available for response. Thirteen patients (48%) met criteria of response (greater than 50% reduction in volume of measurable tumor). Significant myelosuppression resulted from this therapy. Median leukopenia nadir was 3.8 X 10(3) cells/mm3 and median thrombocytopenia nadir was 116 X 10(3) cells/mm3. Additional toxic reactions included anemia, lassitude, anorexia, peripheral neuropath fever, and skin rash. Despite significant, but manageable, toxicity, this combination appears to represent an improvement in the chemotherapy of a traditionaly refractory solid tumor.


Simultaneous assays of cancer-associated antigens in various neoplastic disorders. Five tumor markers were measured simultaneously in serum by radioimmunoassay: carcinoembryonic antigen (CEA), alpha-fetoprotein (AFP), human chorionic gonadotrophin (HGC), the beta subunit of HCG, and Kappa casein. In a population of 935 normal subjects these antigens were undetectable or found within precise limits. In patients with tumors of various origins the rate of pathologically elevated levels was 72% at the beginning of the clinical course (194 cases). This high rate was primarily due to the simultaneous measurement of CEA, betaHCG, HCG, and casein. AFP was of little importance. The simultaneous measurement of these tumor markers may be one biochemical element of diagnosis of carcinoma, although this criterion is neither absolute nor specific, as 14.7% of patients with non-neoplastic disorders (234 cases) were positive for one antigen. In the presence of metastases (112 cases) the rate of pathologic levels of at least one antigen was increased: 86% due to CEA and casein assay at the same time as their absolute levels were increased. Surgical removal reduces the rate of positivity of these antigens to 37%. As was shown in patients with breast cancer, the rate was 10% when the tumor had been removed at Stage N- and 54% when it was removed at Stage N+. Thus, the persistence of pathologic levels could be correlated with the capacity for recurrence or metastases. Finally chemotherapy, radiotherapy, or both, do not decrease the rate of positivity of the tumor markers.


Carcinoid of the uterine cervix: additional observations on a new tumor entity. Twelve carcinoid tumors of the uterine cervix were studied. Based on the microscopic structure, they were divided into well-differentiated and poorly differentiated types. Both tumor varieties affected adult women. The clinical and gross features of these tumors were indistinguishable from those of invasive squamous cell carcinoma. Of the 12 patients, six died, four with poorly differentiated carcinoid and two with the well-differentiated type. Three patients are alive and symptom-free 9 months to 2 years after treatment; the remaining three were lost to follow-up. Microscopically, four well-differentiated carcinoids showed argyrophil cells, and ultrastructurally two had neurosecretory granules. One of the well-differentiated carcinoids resembled islet cell carcinoma and two had areas resembling medullary carcinoma of the thyroid. The poorly differentiated type of carcinoid had a histologic structure and a cell population very similar to that of the oat cell carcinoma of the lung. Of this group, four were argyrophilic, and by electron microscopy four contained pleomorphic secretory granules. It is considered that these tumors arise from the normal argyrophil cell of the cervix and are part of the group of neoplasms of the diffuse endocrine cell system (APUD).


Morphologic and biologic characteristics of adenoid cystic carcinoma cells of the salivary gland. Adenoid cystic carcinoma cells cultivated in monolayer and sponge matrix culture, or implanted on the chorioallantoic membrane (CAM) of embryonated eggs, were observed morphologically, and the glycosaminoglycan components in the tumor tissue were analyzed. This tumor tissue contained a large amount of glycosaminoglycans, composed of chondroitin 4- and 6-sulfate, heparan sulfate, hyaluronic acid, and a small amount of dermatan sulfate. In monolayer culture spindle cells proliferated vigorously as multilayer, and secreated mucinous material. In sponge matrix culture, the proliferating cells became embedded in the material produced by the cells themselves. A trace of fine fibers stained with orceine was observed in the intercellular material in culture. Histologic sections of the implants grown on CAM showed that the tumor cells arranged in various structures produced a large amount of mucinous material that spread into the stromal area without any contribution from the mesenchymal element. The morphologic and biologic characteristics of these tumor cells are quite similar to those of pleomorphic adenoma.


Endodermal sinus tumor of the ovary: a clinical and pathologic analysis of 71 cases. The clinical and pathologic features of 71 endodermal sinus tumors of the ovary were studied in an effort to delineate the histogenesis and biologic behavior of this neoplasm and to evaluate the efficacy of different forms of treatment. Alpha-fetoprotein (AFP) was identified in hyaline droplets, cell cytoplasm, and intercellular spaces of all 15 tumors examined by an immunoperoxidase technique; this supports the view that the neoplasm simulates yolk sac endoderm. There were only nine survivors among 65 patients on whom follow-up information was available; the actuarial survival was 13% at 3 years. Of the neoplasms that recurred, 93% did so within 1 year, and of those patients who died, 93% did so within 2 years. The size and stage of the tumor had prognostic significance, but the patient's age, the mitotic activity, and histologic pattern did not. Although 71% of the patients had Stage I tumors at the time of diagnosis, subclinical metastasis was present in 84% of Stage I patients. Triple chemotherapy (vincristine, actinomycin D, and cyclophosphamide (VAC)) employed after unilateral salpingo-oophorectomy in four patients with Stage I tumors resultivors among 12 Stage I patients treated with combined surgery and radiation. The finding of AFP in all tumors in which this was evaluated suggests that serum radioimmunoassay might be useful to monitor response to therapy.


Embryonal carcinoma of the ovary: a clinicopathologic entity distinct from endodermal sinus tumor resembling embryonal carcinoma of the adult testis. The clinical and pathologic features of 15 examples of a hitherto undescribed germ cell tumor of the ovary are delineated. This tumor resembles the embryonal carcinoma of the adult testis and may be distinguished from the endodermal sinus tumor on the basis of its histologic and immunohistochemical characteristics. An indirect immunoperoxidase method for the localization of human chorionic gonadotropin (HCG) and alpha-fetoprotein (AFP) was done on formalin-fixed paraffin-embedded tissue from 10 neoplasms; HCG was present in all 10 neoplasms, and AFP was found in seven. HCG was indentified only in cells resembling syncytiotrophoblast, whereas AFP was present only in mononuclear embryonal cells, indicating that the two protein antigens were localized in different cell types. Abnormal hormonal manifestations, consisting of precocious puberty, irregular bleeding, amenorrhea, or hirsutism, were present in nine (60%) of the patients. The actuarial survival for the entire group was 39%; for those with stage I tumors, 50%. We are proposing the term "embryonal carcinoma" for this neoplasm in order to distinuish it from the more common endodermal sinus tumor of the ovary and to emphasize the histologic similarity to embryonal carcinoma of the adult testis.