[Study of histamine in asthma].

[Study of histamine in asthma]. Histamine is generally considered as the principal chemical mediator in experimental allergy of type I and anaphylaxis. However, its part in human bronchial asthma remains discussed, because neither the dosage of histamine in blood, nor the use of antihistaminics have furnished any real evidence. This study is dealing with two elements: the importance of local bronchopulmonary phenomena, and the associated mediators also liberated in asthma, such as SRSA. ECFA, kinines, prostaglandins and disorders of balance between, on one hand, alpha and beta-actions of the catecholamines, and on the other hand, between the two main types of cyclic nucleotides. These considerations should allow a better approach of the asthma etiopathology and perhaps an improvement of its treatment.


Splenectomy for palliation of chronic myelocytic leukemia. Sixty patients with chronic myelocytic leukemia (CML) (most, in the "terminal phase" of the disease) were subjected to splenectomy because of symptomatic splenomegaly, thrombocytopenia or anemia for which they required frequent transfusions. Surgical morbidity and mortality were high when the procedure was performed on a "casual" basis, but both were reduced sharply after care of these patients was restricted to a single medical-surgical-nursing team and improved technics of surgery and perioperative management were developed. Significant hematologic and clinical benefit was achieved in half of the patients and temporary arrest of the disease was often observed, but in most patients, the basic evolution of CML was not greatly altered. In eight patients, however, long-lasting improvement (one to nine years) was recorded. Measurement of the doubling time of circulating leukemic cells and other observations were consistent with the hypothesis that, in some patients, the spleen contains a more rapidly proliferating and "more malignant" population of leukemic cells than the marrow. We conclude that splenectomy is often a useful palliative procedure in advanced stages of CML, and that it may be strikingly beneficial in 10 to 15 per cent of such cases.


Technical procedures and staining methods for renal needle biopsies. Procedures specifically developed for kidney needle biopsies are presented. These procedures may be applied to improve the slide quality of all small specimens. Fixation, processing, knife sharpening, cutting, and staining procedures used by the author are discussed. A modification of the Jones method for basement membranes is given.


A light microscopic, autoradiographic study of axoplasmic transport in the normal rhesus optic nerve head. A systematic study of axoplasmic transport in the normal optic nerve head is essential to the understanding of the role of this process in the pathogenesis of disease of the optic nerve. Twenty-one eyes from normal adult rhesus monkeys were subjected to intravitreal injections of tritiated leucine (3H) or proline (3H). Light microscopic autoradiographs were prepared on these eyes and optic nerves six hours to 60 days after injection. The isotope that was incorporated in cytoplasmic components associated with the slow phase of axoplasmic transport (3H leucine) became concentrated in the temporal region of the optic nerve head, unevenly distributed within axons in the axonal bundles, and apparently accumulated in axons or glial cells, or both, in the lamina choroidalis and scleralis as a broad pulse of isotope moved across the optic nerve head. Several anatomic factors, including the distribution of macular fibers, increasing amounts of glial cells within the same axonal bundles as the optic nerve passed from the lamina retinalis to the lamina scleralis, and the abrupt addition of myelin sheaths to axons behind the lamina scleralis, contributed to the complex distribution of isotope in the normal optic nerve head. Glial cell labeling was especially prominent with 3H proline.


Deposition of substances within hydrophilic lenses. One class of hydrophilic lens deposits appears as small, discreted spots under low magnification. Higher magnification of cross sections reveals partially crystalline structures which actually extend into the lens matrix. Histochemical stains indicate the presence of tear film components within these deposits.


Gel-filtered human platelets. Ultrastructure, function, and role of proteins in inhibition of aggregation by aspirin. Gel filtration of human platelet-rich plasma (PRP) on columns of Sepharose 2B removed at least 99.85% of the plasma proteins from platelets when a column 10 cm in height was used and a plasma volume 11 to 14% of the gel-bed volume was applied. ADP and ATP levels in gel-filtered platelets (GFP) were not significantly different from those in PRP. By transmission electron microscopy, GFP were indistinguishable from PRP. Gel filtration appears to be a highly satisfactory technique of separating platelets from plasma without modifying structure, function, or contents significantly. The roles of several crude protein fractions in platelet aggregation and aspirin's inhibition of aggregation were examined. Fraction I (mostly fibrinogen) enhanced collagen-induced aggregation of gel-filtered platelets; Fraction V (mostly albumin) was inhibitory. Fraction II (mostly gamma-globulin) or gelatin had no significant effect. Aspirin added to gel-filtered platelets inhibited aggregation by 80%. The addition of mixtures of plasma proteins containing albumin increased albumin's inhibitory effect. Incubation of gel-filtered platelets with aspirin labeled in the carboxyl position resulted in no uptake of the label. In contrast, incubation with acetyl-labeled aspirin was followed by uptake of more than 2 X 10(6) acetyl groups per platelet in 1 minute. Incubation for 30 minutes resulted in a five- to sixfold further increase in uptake of the label. Aspirin can acetylate platelets and inhibit aggregation directly. Plasma proteins, in particular albumin or a contaminant of the albumin fraction tested, enhance the inhibitory effect of aspirin on platelet aggregation.


Streptozotocin and renal amyloidosis in the Syrian hamster. In the Syrian hamster, administration of the drug streptozotocin was associated with an increased incidence and severity of renal amyloidosis. Some of the more severely affected animals showed a frankly nephrotic picture. Although some of the diseased animals showed decreased levels of serum albumin, no other definite changes in serum proteins were noted, nor was there any evidence of glomerular immunoglobulin or immune complex deposition. None of the lesions previously found after administration of streptozotocin in the Chinese hamster were encountered.


Repetitive extrasystole as an index of vulnerability to ventricular fibrillation. The assessment of ventricular vulnerability by inducing ventricular fibrillation (VF) presents limitations when neural activity is being investigated, especially in the unanesthetized animal. As repetitive extrasystoles (RE) have been observed to precede the occurrence of VF, it was relevant to determine whether the RE threshold provides a reliable index of cardiac susceptivility to fibrillation. The RE and VF threshold relationships were studied in 32 chloralose-anesthetized dogs during left stellate ganglion stimulation, vagus nerve stimulation, and beta-adrenergic blockage with practolol. The vulnerable period was scanned at 1-ms intervals and at 2-mA increments with a single, 2-ms, constant-current cathodal stimulus; RE and multiple RE were induced reproducibly when 66% and 82%, respectively, of the fibrillatory current was administered. The nadirs for RE and multiple RE were coincident in the cardiac cycle with the vulnerable-period threshold for VF. Stellate and vagal stimulation and beta-adrenergic blockade resulted in comparable changes in RE and VF thresholds and produced equivalent shifts in the cardiac cycle of the RE and VF vulnerable-period nadirs. These observations suggest that RE and VF phenomena share a common electrophysiologic basis and that the RE threshold can be used as an end point for measuring ventricular vulnerability to VF.


External diversion for palliative treatment of malignant tracheoesophageal or bronchoesophageal fistulas. The creation of cervical esophagostomy, distal esophageal ligation, and a feeding gastrostomy in combination with an external bypass device for the treatment of malignant tracheoesophageal and bronchoesophageal fistulas offers a simple and rational approach for a critical situation in which more extensive surgical procedures are neither warranted nor more effective.


Pancuronium binding by serum proteins.

Pancuronium binding by serum proteins. Pancuronium has not been suspected of binding to serum proteins. Experimental evidence is presented of strong binding to gamma globulin, and of moderate binding to albumin. No evidence could be found of binding to alpha of beta globulin fractions. Less than 13% of a dose in the clinical range would be unbound and active.


A new look at type I immediate hypersensitivity immune reactions. The pathogenesis of type I disorders, particularly allergic asthma and urticaria, requires a critical review in the light of recent findings which indicate a second pathway for the release of granulocyte mediators. The isolation of enzyme activity of some of these released mediator establishes a direct link between the immunological system and bradykinin, as well as a system of self modulation of kinin activity. These findings open the door for a broader understanding of the asthmatic process and lay the foundation for integrating extrinsic and intrinsic asthma.


Studies of blood clotting system during anaesthesia for prostatectomy. I. Epidural analgesia. The studies were carried out on 20 patients operated on for prostatic adenoma under epidural analgesia. It was observed that activation of the fibrinolytic system is due to trauma to the prostate and is connected with release of plasmin activators into circulating blood. Epidural analgesia has no effect on the blood clotting and fibrinolysis systems and is a safe method in prostatectomy.


10 years' experience treating pancreatic and periampullary cancer. The records of 136 patients with periampullary and pancreatic carcinoma were reviewed and the information compared with other reported series. The clinical presentation with jaundice without other manifestations is associated with the greatest number of potentially curable tumors. The majority of patients were treated by palliative bypass or had exploration and biopsy only. A tissue diagnosis is not imperative before radical excision, providing a systematic preoperative and operative evaluation indicates tumor. Ligation of the pancreatic duct with external drainage results in low morbidity and mortality and good functional results. Radical pancreaticoduodenectomy done in 21 per cent of our patients offers the best palliation and the only hope for cure.


[Changes in the positive P.A.S. reaction of the cytoplasm of giant cells. Role of etiologic factors and the giant cell reaction]. The search for alpha-amylase resistant, P.A.S. positive cytoplasm has been carried out in 206 giant cell lesions with or without specific inflammatory features and with neogenetic and degenerative plasmodia. It allowed to distinguish: --P.A.S. positivity in inflammatory cells of any nature, however with quantitative variations linked to etiologic factors (pathogen agent and disposition), to site, to the age of the plasmodia, and particularly, with negativation of the reaction in ancient lesions;--negativity of the neogenetic and degenerative plasmodia, save the giant cell articular lesions. Cells containing glycogen particularly (muscular tumors, renal or placental) are easily identified thanks to enzymatic digestion tests. Variations observed in the inflammatory cells seem to be the reflect of an active metabolism bringing about resorption phenomena but probably also immunization processes acting at the level of the cellular microenvironment.


Arrhythmias in the mitral valve prolapse syndrome. Prevalence, nature, and frequency. Ambulatory monitoring and maximal treadmill exercise were compared in 40 normal subjects and 31 patients with mitral prolapse. A variable arrhythmia spectrum was observed in prolapse during monitoring: premature ventricular contractions in 18 (58%), supraventricular arrhythmias in 11 (35%), and bradyarrhythmias in 9 (29%). Significantly less arrhythmias occurred in normal subjects during monitoring: 10 (25%, P greater than 0.001), 3 (8%, P less than 0.001), 4 (10%, P less than 0.05), and 2 (5%, P less than 0.02), respectively. In patients with prolapse, arrhythmias occurred on resting electrocardiogram (ECG), 35% premature ventricular contractons, 6% supraventricular arrhythmias, and 10% bradyarrhythmias, and on treadmill exercise, 45%, 10%, and 3%; therefore, ambulatory monitoring was the most sensitive method of arrhythmia detection. No correlation existed between clinical features of prolapse and arrhythmias. Thus, arrhythmias occur in most patients with mitral prolapse, are not predictable by clinical characteristics, comprise a spectrum of ventricular and supraventricular tachyarrhythmias and bradyarrhythmias, and are best detected by ambulatory ECG monitoring.


[Morphometric analysis of periosteocytic osteolysis: its application to the diagnosis of hyperparathyroidism]. The authors indicate the required methodologic conditions for the morphometric analysis of periosteocytic osteolysis, a reliable and specific feature of overactivity of the parathyroid gland. Indeed, significant periosteocytic enlargement has been found in 101 113 cases of primary hyperparathyroidism compared to 92 reference cases. The measurement of the size of the periosteocytic lacunae may be carried out either on hand of a micrometric eyepiece or of a picture analyser. It has to be carried out on decalcified bone sections of 5 microns thickness and it is necessary to measure at least 50 lacunae. The decalcification process unmasks an already partially decalcified crown of periosteocytic bone tissue the thickness of which is more important in the hyperparathyroidal bone than in normal bone. This zone has a peculiar collagen texture. The measurement of the peri-osteocytic enlargement is easy on stained sections of decalcified bone or on microradiographs of non-decalcified bone and, contrariwise, very dubious on stained sections of non-decalcified bone.


Diagnosis of allergy to stinging insects by skin testing with Hymenoptera venoms. SKin testing was done on 30 patients with a history of anaphylactic reactions after a Hymenoptera sting and on 30 control subjects. The patients all had positive basophilhistamine release to one or more venoms on challenge with the specific venoms used for skin testing (honey bee, yellow jacket, white-faced hornet, yellow hornet, and Polistes). At 0.1 mug of venom/ml and at 1.0 mug of venom/ml, 75% and 100%, respectively, of the sensitive patients had a positive skin test. There was a significant (P less than 0.001) correlation between skin test and histamine release results. Of the 150 skin tests in control patients, only 1 was positive. Venom skin tests provide, for the first time, a simple, readily available technique to accurately diagnose allergy to stinging insects.


[Alpha-fetoprotein. Significance and current importance]. The identification of AFP in the serum after birth is always due to a hepatoma or, more rarely, a teratocarcinoma. The increasing sensitivity of methods of immuno-chemical investigation and radioimmunoassay, permit today its identification in cases other than hepatoma. In the foetus, the hepatocyte or its precursor is the site of main synthesis but is not exclusive. The synthesis of AFP has been demonstrated both in the cells of endodermic origin, yolk sack, anterior part of the intestine, stomach and colon. In hepatomas, only a small proportion of neo-hepatocytes is AFP positive on immunofluorescence or on electron microscopy. A quantitative study of AFP is useful above all: a) During pregnancy: detection of foetal distress or malformation when AFP is present in the amniotic fluid. b) For the supervision of patients after surgical treatment. The interest of AFP detection and estimation using high sensitivity methods is emphasized.


Biochemical basis of the serological cross-reactions between Brucella abortus and Yersinia enterocolitica serotype O:9. A method based on the inhibition of agglutination is described that may be used for the differential serological diagnosis between B. abortus and Y. enterocolitica serotype O:9. An antigen with high immunological capacity was isolated from Brucella. This antigen inhibited both homologous and heterologous agglutination by Brucella antiserum, but only the heterologous agglutination by Yersinia antiserum. It proved to be constitued of a polysaccharide (N-acetylglucosamine, glucose, mannose and 2-keto-3-deoxyoctonic acid), a protein and a phosphoglycerid moiety. Lipid A was absent from the Brucella antigen. Incomplete polysaccharide synthesis of the Brucella antigen, with concomitant loss of serological specificity by the rough mutant has been described. Oligosaccharides containing N-acetylgalactosamine, glucose and galactose were isolated from the specific side chain of Yersinia lipopolysaccharide. Lipid A constituents were also identified in the latter.


[Alpha-fetoprotein and neonatology: Postnatal estimation of gestational age and idiopathic respiratory distress syndrome (author's transl)]. Cord serum AFP levels have been determined in 66 newborn samples, between 30 to 42 weeks of gestational age. Results showed some relation between AFP rate and newborn maturity degree, so it was possible to foresay the gestational age with some limits of error +/- 17 days in full-term newborn, and +/- 10 days in premature ones. Cuantification in the amniotic fluid of 18 high risk gestation gave normal AFP rates. AFP of thirty patients with respiratory distress was tested. An AFP increase was present in 14 of 15 cases of hyaline membrane. On the remaining 15 cases of respiratory distress only 3 gave high levels. These facts point that AFP can be used as a suitable parameter in the biochemical diagnosis of hyaline membrane.


Characteristics of antigenic and biological properties of influenza A viruses isolated different epidemics of the same cycle. The antigenic character of the haemagglutinin and of the neuraminidase from Myxovirus influenzae strains isolated in the USSR during the 1968, 1970, 1971-1972 and 1972-1973 epidemics, are studied. The haemagglutinins were, at first, relatively, homogenous and constant, but subsequently changed gradually newer isolates became less and less reactive against a specific serum prepared with A/Hong Kong/68. However, this druft up to 1972-1973, did go beyond the limits of the H3 subtype. There were also some differences in the biological properties between the earlier and later haemagglutinins, especially with respect to their sensitivity to gamma-inhibitor in animal sera, and to their pattern of thermoinactivation at 56 degrees C. There was a similar antigenic drift in the case of the neuraminidase, but here it was more pronounced than with the haemagglutinin, for the 1972-1973 isolates had lost virtually all cross reactivity against the 1957-1969 strains. In this manner, three neuraminidase variants could be distinguished: N2-1 from H2 sub-type isolates since 1957, N2-2 from 1964-1971 isolates and N2-3 from 1972, and beyond, strains. Central strains of virus, such as England/42/72 and some of the Leningrad/72, had neuraminidases intermediary between N2-2 and N2-3. The in vitro thermostability of these neuraminidases was variable, according to the strains. The explosive nature of the 1972-1973 epidemic was no doubt linked to the changes in the virus neuraminidase, which were more pronounced than the changes in the haemagglutinin.


Immunological mechanisms and diagnostic tests in allergic drug reactions. Current knowledge about the immunological mechanisms involved in drug hypersensitivity is reviewed, with comments on the major conditions which favour the development of reactions to drugs. In spite of the numerous tests available to establish the in vitro diagnosis of allergic reactions to drugs, it is often very difficult to relate the clinical picture to the laboratory findings. The results of various tests of antibody and cell-mediated responses are presented and discussed.


[Action of aurantin on the chromosomal structure of continuous cultures of human amniotic cells (line A-1)]. When transplantable cultures of human amniotic cells, line A-1 were treated with auranthin in concentrations of 0.1 and 0.5 gamma/ml for 48 hours of their growth, the chromosomes in the methaphase proved to be not evently spiralized longitudinally and acquired the appearance of segmented structures. All the chromosomes in the set possessed the capacity for such segmentation. It was most pronounced near the centromers and at the ends of the meta- and submetacentric chromosomes. The acrocentric chromosomes were segmented longitudinally more evenly. Loose segments of the chromosomes most probably corresponded to the areas, in which auranthin binding with DNA occurred. The effect of auranthin on differentiation of the chromosomes longitudinally may be used, as it was indicated for actnomycin D, in identification of homological chromosomes in the karyotype, as well as in studies on the temporal sequence of DNA synthesis in the chromosomes.


[Effect of phytobacteriomycin, trichotecin, hygromycin B and levoristatin on some rat liver enzymes]. In experiments with albino rats it was found that after administration of phytobacteriomycin, trichotecin, hygromycin B or levoristatin into the stomach in doses of 1/20 of LD50 activity of the microsomal enzymes of the liver cells significantly changed and the changes persisted within at least 2 weeks. The above antibiotics induced similar changes in the lysosome enzyme, i.e. acid phosphatase, providing an increase in its activity. Changes in the activity of succinate dehydrogenase (mytochondria indicator enzyme), glucose-6-phosphatase (ribosome indicator enzyme) and aspartate aminotransferase (cytoplasm indicator enzyme) were different for each antibiotic. It is concluded that the above antibiotics were capable of impairing on intoxication the enzymatic function of various cell microstructures, though the levels of the change direction may be different.


Phenylalanine stimulation of gramicidin S formation. Bacillus brevis produces the antibiotic gramicidin S, a cyclic peptide comprised of two pentapeptide units of the sequence d-Phe-l-Pro-l-Val-l-Orn-l-Leu. Synthesis of this antibiotic is carried out by two enzymes, GS synthetases I and II. By supplementing a defined minimal medium [glycerol, NH(4) (+), tris(hydroxymethyl)aminomethane, salts] with each of the constituent amino acids (in the l form), we found that only l-phenylalanine had a marked stimulatory effect on gramicidin S production. This effect was not caused by an increase in growth, by an induction of GS synthetases I or II, or by a stabilization of these enzymes. l-Phenylalanine apparently stimulates production via its role as a precursor of the d-phenylalanine moiety of the gramicidin S molecule.


Electron transport components of the MnO2 reductase system and the location of the terminal reductase in a marine Bacillus. The response of MnO2 reduction by uninduced and induced whole cells and cell extracts of Bacillus 29 to several electron transport inhibitors was compared. MnO2 reduction with glucose by uninduced whole cells and cell extracts was strongly inhibited at 0.1 mM dicumarol, 100 mM azide, and 8 mM cyanide but not by atebrine or carbon monoxide, suggesting the involvement of a vitamin K--type quinone and a metalloenzyme in the electron transport chain. MnO2 reduction with ferrocyanide by uninduced cell extracts was inhibited by 5 mM cyanide and 100 mM azide but not by atebrine, dicumarol, or carbon monoxide, suggesting that the metalloenzyme was associated with the terminal oxidase activity. MnO2 reduction with glucose by induced whole cells and cell extracts, was inhibited by 1 mM atebrine, 0.1 mM dicumarol, and 10 mM cyanide but not by antimycin A, 2n-nonyl-4-hydroxyguinoline-N-oxide) (NOQNO), 4,4,4-trifluoro-1-(2-thienyl),1,3-butanedione, or carbon monoxide. Induced cell extract was also inhibited by 100 mM azide, but stimulated by 1 mM and 10 mM azide. Induced whole cells were stimulated by 10 mM and 100 mM azide. These results suggested that electron transport from glucose to MnO2 in induced cells involved such components as flavoprotein, a vitamin K-type quinone, and metalloenzyme. The stimulatory effect of azide on induced cells was explained on the basis of a branching in the terminal part of the electron transport chain, one branch involving a metalloenzyme for the reduction of MnO2 and the other involving a metalloenzyme for the reduction of oxygen. The latter was assumed to be the more azide sensitive. Spectral studies showed the presence of a-, b-, and c-type cytochromes in membrane but not in soluble fractions. Of these cytochromes, only the c type may be involved in electron transport of MnO2, owing to the lack of inhibition by antimycin A or 2n-nonyl-4-hydroxyquinoline-N-oxide. The terminal MnO2 reductase appears to be loosely attached to the cell membrane of Bacillus 29 because of cell fractionation it is found associated with both particulate and soluble fractions. Electron photomicrographs of bacilli attached to synthetic Fe-Mn oxide revealed an intimate contact of the cell walls with the oxide particles.


Palliative surgical treatment of gastric carcinoma, duration of survival in 217 cases. During the period 1958 to 1972 inclusive, 217 patients were subjected to a palliative operation because of carcinoma of the stomach. Subtotal gastric resection was performed in 33 and a total resection in 42 cases. The postoperative mortality in the resected group amounted to 8%. The average duration of survival with exclusion of the postoperative mortality amounted to 17 months with a median duration of 12 months. Five patients in this group (7%) survived for 5 years, and two patients are still alive without recurrence 9 and 6 years, respectively, after the operation. Among the 142 remaining patients, the postoperative mortality was 23%. The average survival with exclusion of the postoperative mortality was 5 months; the median duration of survival amounted to 4 months. The average duration of survival among the patients subjected to a gastro- or jejunostomy was shorter: 3 months, with a median of 2 months. Of the various palliative measures, resection is to be preferred. If it is not feasible, gastro-enterostomy and intubation may be considered. In a limited number of cases, the alimentary fistula may still be indicated, even though it is an unsatisfactory type of palliation. Hoerr's classification is satisfactory as a clinical subdivision. The dividing line between operations with curative and with palliative intent as a rule is put at B III. Of the patients treated palliatively, 92% had died after 2 years; 79% of those subjected to resection and 100% of those subjected to some other intervention.


Surgical management of carcinoma of the junction of the main hepatic ducts. Twenty-six patients are reviewed who had primary carcinomas involving the junction of the hepatic ducts. The majority had had an initial procedure of palliative biliary diversion elsewhere and were referred for further treatment. In three cases, en bloc resection of the tumor with total hepatectomy and orthotopic liver transplantation were performed. All tumor growth was encompassed in each case, but within 4 months all succumbed as a result of allograft rejection. Auxiliary (heterotopic) liver transplantation was performed in another patient because of recurrent disease after previous left hepatic resection in continuity with a hilar duct lesion. Five patients underwent hepatic lobectomy with en bloc resection of the hepatic duct junction. When adequate tumor excision was not feasible, biliary diversion could provide good palliation in some instances for extended periods of time. This is demonstrated by one patient who lived for 4 years and 4 months after the initial operation. In the meantime, the patient underwent 6 subsequent procedures of dilating of constricted bile ducts and tube cannulation of the biliary tree. Biliary diversion was achieved in 4 cases by intrahepatic cholangiojejunostomy. One of these patients, who is on chemotherapy, is asymptomatic one year after surgery.


[Action of injected disopyramide on ventricular arrhythmias and atrioventricular conduction]. The authors have studied the bathmotropic and dromotropic effects of intravenous injection of 100 mg of disopyramide, the therapeutic dose being 1 to 2 mg/kg. Out of the 28 severe arrhythmias studied, it was the ventricular extrasystoles which benefited most from treatment (79% completely successful, while there was a lesser degree of success in treating the ventricular tachycardias (35% completely successful, 30% partially successful). The negative dromotropic effect of injected disopyramide on sub-nodal conduction was studied by an endocavitary technique in 10 patients presenting with a spontaneous onset of disordered intracardiac conduction; the effects were moderate. With the exception of 4 cases who sustained complications involving rhythm and haemodynamics, the clinical and cardiovascular tolerance of patients to the drug appeared to be satisfactory.


Creatine phosphokinase levels in children with severe developmental disturbances. Serum creatine phosphokinase (CPK) levels were studied in individuals: 40 psychotic children suffering from childhood autism, atypical personality development, and childhood schizophrenia; five children with childhood aphasia; 22 children with severe personality disorders; 29 normal children and normal siblings of psychotic children; and 14 normal parents of psychotic children. Creatine phosphokinase levels from the entire population of adults and children were normally disturbed, and the mean CPK levels for the eight diagnostic groups were within normal limits. Those 22 children with personality disorders had significantly higher CPK levels than the other diagnostic groups. This relatively higher level of CPK may be related to vulnerability to later development of schizophrenic spectrum disorders. There was no apparent relationship between CPK levels and motor activity, nor was there any change in the level of CPK during a trial of psychoactive medication. Creatine phosphokinase levels remained relatively stable on test-retest determination.


Changes in the cells of the adenohypophysis associated with the diadromous migration of the threespine stickleback, Gasterosteus aculeatus L. The adenohypophysis of the threespine stickleback, Gasterosteus aculeatus, was studied light-microscopically to determine and estimate the cell types and their function. For these purposes, the adult specimens obtained during the period from migration to spawning were examined. Further, the juveniles caught in the spawning bed were subjected artificially to sea water. The rostral pars distalis (RPD) consists mainly of two types of cells: dorsally shifted lead hematoxylin (PbH)-positive cells bordering the neurohypophysis correspond to corticotrophs, and antero-ventrally shifted acidophil cells are identified as prolactin cells. The latter undergo marked hypertrophy and active state just at the time of entering the river (February), while no detectable change was seen in the former throughout anadromous migration. The role of prolactin on the osmoregulation in freshwater environment is thus suggested. The proximal pars distalis (PPD) consists mainly of two cell types: the basophil cells in round shape are regarded as the gonadotrophs and the acidophil cells in ellipsoid shape are considered to be somatotrophs. The size of the gonadotrophs reaches the maximum at the time of spawning. A few AF-positive cells of elongate shape occur in the dorsal region and are identified as thyrotrophs. In the pars intermedia (PI), two types of cells are discernible: PAS-positive and PAS-negative cells. The latter attained their maximal size in the earliest time of anadromous migration.


[The cytobacteriological coloring substance BF (methylene blue and basic fuchsin)]. The Authors have set up a cytobacteriological colouring matter for the reading of the figured elements of liquor, urine, exudats, and trasudats. The colouring solution is steady, it colours and fixes at the same time and is composed by blue methylene and basic fuchsine. There is abready a combination of both colouring matters in WAYSON'S formula for the bipolar colouring matter of the Pasteurella pestis and in PICK'S and Jacobson's formula to point out the intercellular neisserie.


HLA typing.

HLA typing. I present an overview of HLA typing, a method of evaluating donor-recipient compatibility. The major histocompatibility system in man, the HLA region, considered to control the immunologic response to transplantation and perhaps to certain diseases, is discussed in relation to chromosomal location, chromosomal mapping, association with other markers, nomenclature of the gene products (the HLA antigens), inheritance of the HLA antigens, and methods of determining these antigens. Use of HLA typing is currently most applicable in the correlation of certain diseases with HLA antigen frequency, in evaluating donor-recipient pairs for renal and bone marrow transplantation and for transfusion of platelets and granulocytes and in paternity testing.


'Acidophilic' pituitary tumors: a reappraisal with differential staining and immunocytochemical techniques. The cells of pituitary adenomas classified as acidophilic on PAS-light green-orange G staining could be further differentiated with the Brookes technique: they stained nonspecifically bluish-gray, orange with orange G, or red with carmoisine. On immunostaining for growth hormone and prolactin, the gray cells were either negative or reactive for prolactin, the orange cells contained growth hormone, and the red cells contained prolactin. Of 28 tumors, eight showed no immunostaining, 11 stained only for prolactin, three stained only for growth hormone, five contained mostly growth hormone cells and some prolactin cells, and one contained predominantly prolactin cells but also numerous growth hormone cells. Immunoreactive growth hormone granules in adenoma cells were usually arranged randomly; prolactin granules were often concentrated along one nuclear pole. This study emphasizes the tinctorial and immunocytochemical heterogeneity of "acidophilic" adenomas.


Prostatectomy in an 85-year-old hemophiliac. Improved factor VIII preparations have made major surgery feasible for hemophiliac patients, as well as increasing their survival. In a hemphoiliac undergoing prostatectomy, the effects of local or systemic fibrinolysis and possible disseminated intravascular coagulation must be considered, in addition to the factor VIII deficiency. We successfully treated an octogenarian with benign prostatic hypertrophy and mild hemophilia during and after suprapubic prostatectomy by infusions of antihemophilic factor (factor VIII) concentrate. Cessation of infusion on the 15th day resulted in bleeding two days later. This ceased after resumption of antihemophilic factor infusion. Tests for abnormal fibrinolysis and fibrin split products gave negative results.


[Modification of Bowie's method of demonstrating specific granules in cells of the human renal juxtaglomerular apparatus fixed in neutral formalin]. A modification of Bowie's method for detection of specific granules of the juxtaglomerular apparatus of the human kidneys, fixed in 10% neutral formalin, is suggested. In order to achieve better staining, sections of material fixed in formalin are additionally treated with Helly's liquid and, following the removal of sublimate deposit, with a 2.5% solution of potassium bichromate. After this the sections are stained by Bowie's method in accordance with Pitcock and Hartroft's prescription.


Bilateral amyloidosis of the vitreous body: report of a case without systemic of familial involvement. Amyloidosis of the vitreous body is an uncommon and often misdiagnosed condition that causes progressive visual loss. It is usually associated with primary familial systemic amyloidosis. The patient described in this report has amyloidosis of both vitreous bodies; her case is unusual because no other family members are affected and because there has been no evidence of systemic involvement over a seven-year observation period. After part of the vitreous body of her right eye was removed surgically, visual acuity improved from light perception to 20/60. New concepts regarding the nature, classification, and histochemical identification of amyloid are discussed.


Antigenic structures of Salmonella flagella. I. Presence of an antigenic determinant exposed at one end of flagellar fragments. Salmonella flagellin, which is a constitutional subunit of the flagellum, was shown to have antigenic determinants distinct from its own serotypic ones. These antigenic determinants were found to be common to flagellins from the so-called g-complex serotypes, such as fg, mt, gm, gt, gp and gmptu, but not to those from other serotypes, such as a, i or enx. Rabbits immunized with flagellin of serotype "fg" produced anti-"fg" flagellin antibodies. Only about 20 percent of these corresponded to the serotype determinants of the "fg" on the surface of the flagella, and the remaining 80 percent reacted with the flagellin of the unrelated serotype "mt", and corresponded to the distinct determinants common to the flagellin molecules. These antigenic determinants were detected by the immunoferritin technique at only one, not both, terminals of the flagellar fragments, suggesting that a unidirectional arrangement of flagellin subunits in the flagella may expose the inherent conformation of the subunits at only one end of the flagellum.


The mechanism of sensitivity to phleomycin in growing Escherichia coli cells. Stationary-phase Escherichia coli B cells transferred to new growth medium are initially resistant to net DNA breakage by low concentrations of phleomycin, and become sensitive as DNA replication commences. From studies with inhibitors of various stages of the DNA replication cycle it is evident that it is not DNA synthesis itself that is required for induction of DNA breakage by phleomycin, but events associated with the initiation of DNA replication. Termination of replication in the absence of further initiaiton results in resistance to phleomycin. The cellular change responsible for changes in sensitivity to phleomycin could be the attachment of the bacterial chromosome to the cell membrane at initiation and detachment on termination of replication, suggesting an alteration in the balance between cellular DNA breakage and repair processes for membrane-associated compared with non-membrane-associated DNA.


[Adams-Stokes syndrome in a case of Mobitz type II AV-block triggered by premature atrial contractions]. The case of a patient with Mobitz type II AV-block is presented who suffered from recurrent dizzy spells and syncopal attacks. These episodes were due to intermittent asystoles lasting for 3-17 seconds, and it could be shown that they were triggered by two or more successive premature atrial contractions. The observation that there were no subsidiary escape beats or rhythms during the asystolic intervals and the ECG pattern for the conducted beats (RBBB and LAH) suggest an intraventricular (trifascicular) level of AV-block. The exact analysis of the asystolic pauses makes it likely that these were initiated by the penetration of the premature atrial impulses into the left posterior subdivision of the left bundle (concealed conduction). The present case demonstrates the fact that premature atrial contractions may produce prolonged asystolic attacks in patients with advanced intraventricular conduction disturbances.


[Linear- and Areal-analytical results of the karyometric studies on monolayer cultures and the influence of actinomycin (author's transl)]. Actinomycin D (AMD) inhibits the RNA-synthesis by intercalating with the DNA-helix. In monolayer culture of embryonic mouse brain cells in the 8th passage the cell nuclei were demonstrated with the Feulgen-reaction and quantitatively-morphometrically evaluated following a 48 h intoxication with AMD (0.001; 0.01; 0.1 mug/ml). The mean value of nuclear area calculated from single measurements with the Quantimet 720 is significantly larger when compared to the controls at a dose of 0.01 mug AMD/ml. At 0.1 mug AMD/ml ca. 30% of the cell nuclei are pyknotic; the non-pyknotic nuclei are enlarged significantly by appr. 20%. The linear-analytic results using a Digiscan show no enlargement of tendon length, on the contrary, at 0.1 mug AMD/ml a significant diminution versus the controls can be seen. This resutls from the pyknotic cell nuclei which are also measured in this case no single measurements were carried out. The determination of cell density using a Digiscan shows a diminution of cell number by 19, 31 or 68% dependent upon AMD-concentration.


Inhibition of denaturation of human gamma globulin by a mixture of L-histidine, L-cystine, and copper, and its clinical implication in rheumatoid arthritis. A mixture of histidine, cystine, and copper mimicked gold thiomalate, N-ethylmaleimide, and p-chloro-mercuribenzoic acid in inhibiting sulfhydryl-disulfide interchange-mediated denaturation of human gamma globulin, bovine serum albumin, and diluted human serum. Measurable inhibitory effects were obtained with a mixture of physiologic concentrations of L-histidine, L-cystine, and copper. This work suggests a mechanism by which the hypohistidinemia of rheumatoid arthritis could contribute to the pathogenesis of the disease.


Purification and translation of an immunoglobulin lambda chain messenger RNA from mouse myeloma. Here we describe the 500-fold purification of an mRNA encoding an immunoglobulin lambda light chain derived from the mouse myeloma tumor, RPC-20. Purification involves the isolation of membrane-bound polysomes, oligo(dT)-cellulose chromatography, and sucrose gradient centrifugation under conditions favoring denaturation of polynucleotide complexes. The mRNA purified in this way directs the cell-free synthesis of a polypeptide which is five or six amino acids longer than the mature form of RPC-20 light chain. In addition to directing the synthesis of a precursor-like polypeptide, the mRNA migrates on electrophoresis as a band containing approximately 1150 nucleotides, about 500 more than required to encode the mature form of the light chain.


Rheumatoid factor (antigammaglobulin) in women: effects of oral contraceptives use of its prevalence. A total of 14,856 women, including 921 pregnant subjects, were tested for rheumatoid factor; 4,562 were using oral contraceptives at the time of testing. The prevalence of rheumatoid factor increased directly with age. The age-adjusted prevalence of rheumatoid factor was lower in oral contraceptive users than in nonusers but this difference was not statistically significant. Rheumatoid factor remained positive in 39% of subjects undergoing retesting after an average interval of 16 months. Those women with higher titers of rheumatoid factor were more likely to remain positive (81%). Of the women having positive tests, 5.4% were identified as having rheumatoid disease.


Conformational transitions of non-helical proteins effected by dodecyl sulfate. Circular dichroism of alpha1-acid glycoprotein, Bence Jones protein, carbonic anhydrase B, deoxyribonuclease, pepsinogen, and plasminogen. Circular dichroism (CD) of serum alpha1-acid glycoprotein, urinary Bence Jones protein, human carbonic anhydrase B, deoxyribonuclease from bovine pancreas, porcine pepsinogen, and plasminogen from human serum was tested in the absence and presence of 0.005-0.05 M sodium dodecyl sulfate. It was found that in all cases the CD spectra of these proteins were modified by the dodecyl sulfate into spectra indicating the presence of a moderate content of alpha-helix. The transitions were enhanced by addition of acid (pH 2.1-4.4) in all cases tested. Comparison of the various proteins with respect to the amount of reconstruction of the main chain conformation showed that the amount of helix formed depended on the amino acid composition of the protein. Rigidity due to cross-linking by disulfide bridges is the strongest deterrant to the conformational change of the main chain. The CD bands of the native proteins in the 250-350 nm spectral zone were extinguished by sodium dodecyl sulfate, and new weak bands were observed the positions of which corresponded approximately to those of the native proteins. In all cases, except the carbonic anhydrase B, the bands of thus denatured proteins were negative.


Antibodies to native and denatured collagens in sera of patients with rheumatoid arthritis. A passive hemagglutination assay was used to detect antibodies to native human collagens and to collagen chains in the sera of 110 rheumatoid patients and those of 75 normal controls. The incidence and titer of anticollagen antibodies in rheumatoid arthritis are high, but in controls they are low or in most instances absent. No correlation was found between the stage of RA, or titers of rheumatoid factor, or ANA and the incidence and/or titers of antibody to any given type of collagen.


A kinetic study of the inhibition of human and bovine trypsins and chymotrypsins by the inter-alpha-inhibitor from human plasma. Human plasma inter-alpha-inhibitor forms 1:1 inactive complexes with human and bovine trypsins (EC 3.4.21.4) and chymotrypsins (EC 3.4.21.1). The association and dissociation rate constants as well as the equilibrium dissociation constants (Ki) of the complexes formed of inter-alpha-inhibitor and the four proteases have been measured. The most stable complexes are those formed with the bovine enzymes. For instance, Ki = 2.1-10-11 M for bovine trypsin whereas Ki = 1.2 - 10-8 M for human trypsin. Whatever the species, the complexes formed with the chymotrypsins are less stable than those formed with the trypsins.


[Study of the complex between porcine plasmin and alpha2-macroglobulin (author's transl)]. Porcine plasmin (EC 3.4.21.7) is obtained from plasminogen activated by human urokinase. This enzyme can bind, in an equimolecular ratio, to an alpha2-macroglobulin isolated from porcine serum. The number of active sites of plasmin has been determined through a burst titration of nitroaniline during the presteady-state hydrolysis of an amide substrate (N-alpha-carbobenzoxy-L-arginine-p-nitroanilide). The kinetic constants relative to a very sensitive ester substrate (N-alpha-carbobenzoxy-L-lysine nitrophenylester) hydrolysis have been measured. The binding of plasmin to alpha2-macroglobulin results in a complete inhibition of proteolytic activity, a reduction of active sites number and a decrease of esterolytic activity towards this substrate. In the complex, the residual activity (about 60%) is protected from protein inhibitors. Absorbance difference spectra show that 1 mol of alpha2-macroglobulin binds 1 mol of plasmin and 2 mol of trypsin. When plasmin is first bound to alpha2-macroglobulin, only 1 mol of trypsin can gain access tothe second site without removing the plasmin, showing that a steric hindrance is implicated in the inhibition performed by alpha2-macroglobulin binding.


Activation of fibroblast procollagenase by mast cell proteases. Proteases capable of activating procollagenase from gingiva and from fibroblast and macrophage monolayer cultures were harvested from homogenates of canine tumor mast cells. The mast cell proteases lysed casein and Azocoll but not native collagen. In low salt concentrations the enzymes existed at high molecular weight complexes, which were dissociated by increasing the salt concentration above 1.0 M (NaCl, KCl). Gel filtration in 1.4 M KCl separated the protease activity into three peaks, all of which activated procollagenase. Two of the enzymes showed substrate specificities (hydrolysis of p-tosyl-L-arginine methyl ester and benzoyl-tyrosine ethyl ester) and reactive center reactivities similar to pancreatic trypsin and chymotrypsin. Based on gel filtration, apparent molecular weights of 160 000 (p-tosyl-L-arginine methyl ester esterase), 90 000 (main procollagenase activator) and 36 000 benzoyl-tyrosine ethyl ester esterase) were determined. Activation of procollagenase resulted in a 18-20 000 decrease of the molecular weight. The activation was directly related to the amount of activator added within certain limits. Further addition of activator resulted in proteolytic inactivation of collagenase.


Comparison of fungal glucose oxidases. Chemical, physicochemical and immunological studies. Glucose oxidases (beta-D-glucose:oxygen 1-oxidoreductase, EC 1.1.3.4) from two fungal genera (Aspergillus and Penicillium) were studied chemically, physicochemically and immunologically to elucidate the similarities and dissimilarities between these enzymes. Investigation of circular dichroism spectra revealed that these enzymes proteins possess essentially identical conformations. However, differences found in thermal inactivation parameters, catalytic parameters and quantitative immunological reactivities indicate that these enzymes must have some minor but distinct variations in their structures. Interestingly, it was observed that the Penicillium enzyme cross-reacted with the antiserum against the Aspergillus enzyme with an association constant of two orders of magnitude lower than that of the Aspergillus enzyme, and that the precipitin one of the Penicillium enzyme fused together with that of the Aspergillus enzyme in the immunodouble diffusion test. These results lead to the conclusion that these enzymes are closely related but not completely identical, and suggest that they might have evolved from a common ancestral precursor.


Temperature-jump-induced release of hydrogen ions from chloroplasts and its relaxation characteristics in the presence of ionophores. Temperature-jump-induced absorption changes of a bromocresol purple in chloroplast suspensions in the dark were studied. After a rapid rise in temperature (less than 10 mus), a slow absorbance decrease of bromocresol purple (t1/2 approximately 0.2 s) following a fast absorbance decrease of chloroplasts and bromocresol purple (t1/2 less than 1 ms) was observed. The slow absorbance decrease corresponds to acidification of the suspending medium, indicating H+ efflux from chloroplasts after the temperature jump. Nigericin and gramicidin D suppressed the slow absorbance change completely in the presence of 10 mM KC1, while valinomycin did not affect it. The fast absorbance change was not affected by the above ionophores. 3-(3,4-dichlorophenyl)-1,1-dimethylurea also diminished the slow absorbance change.


[Autoantibodies to the common antigenic determinant of group A streptococcal polysaccharide and epithelial cells of mammalian thymus and skin]. A cross-reactive (CR) antigenic determinant in the polysaccharide of group  A streptococus and the mammalian thymus and skin was studied. The CR antigen was found in adult humans and human embryos irrespective of the blood group, as well as in the tissues of all the animal species studied, including rabbits immunized with streptococcus and producing antibodies to A polysaccharide. Evidence was obtained that the antibodies elaborated to the CR determinant of polysaccharide A and of the tissue epithelium should be classed as autoantibodies. It is suggested that reaction of these antobodies with the CR antigen in the thymus could serve as one of the causes of autoimmune thymitis during rheumatic fever.


Acute nonlymphocytic leukemia in adults: correlations with Q-banded chromosomes. Chromosome banding patterns were obtained for 50 of 55 consecutive adult patients with acute nonlymphocytic leukemia during a 5-yr period. Twenty-two of the 50 cases were diagnosed as acute myelocytic leukemia (AML), 24 as acute myelomonocytic leukemia (AMMol), 2 as acute promyelocytic leukemia (APL), and 2 as erythroleukemia. Twenty-five patients had initial chromosome abnormalities during the course of the disease. The median survival of patients with normal chromosomes initially (group I) was 10 mo, whereas that of patients with abnormal chromosomes initially (group II) was 2 mo. Similar times were obtained for treated patients with AML and AMMol. However, when the AML patients were separated into those with and those without a chromosome abnormality, the median survival times were markedly different (2 mo versus 18 mo, respectively). Patients with AMMol demonstrated no difference in median survival times when subgrouped according to the presence or absence of chromosome abnormalities. The treated group II patients whose marrow samples had only abnormal metaphases had a poorer response (10% complete remission) and median survival (2 mo) than the group II patients who had at least one normal metaphase (42% complete remission with a median survival of 9 mo). The two cases of APL demonstrated a deletion of the long arm of No. 17 which occurred in the same region of the chromosome in each case. Both patients had similar clinical histories, with disseminated intravascular coagulation, and neither responded to therapy.


Screening for spina bifida cystica. A cost-benefit analysis. The costs and economic benefits are examined of introducing a programme for the mass-screening of pregnancies for the detection and abortion of fetuses with spina bifida cystica. A benefit-cost index is derived, and the possible effects on it of making different input assumptions are discussed. It is considered that, on economic grounds, screening may be worthwhile only in populations in which the incidence of spina bifida is high.


C- and G-bands of the opossum chromosomes: terminal sequences of DNA replication. The sex chromosomes of the opossum, Didelphys virginiana, are the only elements that exhibit C-banding. In contrast, the sex chromosomes as well as the autosomes bear specific G-Bands. However, unlike other mammalian species different types of G-banding are observed if the chromosomes are pretreated with trypsin and SSC solution The SSC-pretreated chromosomes show discrete bands only when stained with Giemsa at certain pH values. An asynchronous pattern of terminal DNA replication is observed among the three C-banding regions of the X-chromosome. The inter- and intrapositive G-banding areas of the chromosomes are not always late in DNA replication in comparison to those negatively stained G-banding areas.


Additional observations on the preparation of R banded human chromosomes with acridine orange. A number of technical factors which affect acridine orange R banding (RFA banding) were studied. These variables included age of slide, timing of fixation, details of incubation mounting and the use of sequential technics. Optimal RFA banding was obtained between 15 and 20 days but good or very good preparations were obtained between 7 days and 2 months. Improved results were obtained in slides that were 3-4 months old by refixing the slides in ethanol acetic acid. Intermittent movement of slides during incubation in buffer as well as the details of mounting and removal of cover slips were found to be important. The best sequential banding was obtained with the sequence of Q to R but good results were obtained with the sequence G to R using ASG banding. Satisfactory results with the sequence R to C were not obtained. With careful attention to these variables good RFA binding can be obtained over a period of several months.


Unresectable squamous cell carcinoma of the lung and its management by combined bleomycin and radiotherapy. A clinical study of the enhanced results. The interaction between the antibiotic bleomycin and x-radiation has been studied in vitro and in vivo. Tissue culture results appear to reflect in vivo sensitivities correctly. Simultaneous exposure to bleomycin and gamma radiation enhances killing of both sensitive and resistant lines. Data from a pilot study combining bleomycin with conventional radiation for unresectable squamous cell carcinoma of the lung suggest that the simultaneous administration of bleomycin (10 mg/m2 intravenously twice weekly) with short-course radiation treatment is well tolerated and without dangerous pulmonary complications. Tumor response was greater in the combined-therapy group (46%) than in radiation-only controls (26%); median survivals were 13 and 6 months, respectively. Unlike previously published data, responders appeared to have a significant survival advantage over nonreponders, suggesting that bleomycin may be slightly effective in inhibiting the development of systemic metastasis, and that it positively enhanced local control of primary disease.


De novo Kaposi's sarcoma in renal transplantation. Case report and brief review. This report describes a de novo development of Kaposi's sarcoma in a Puerto-Rician man 9 months after a cadaveric renal transplant. Progression of the disease was observed despite local irradiation, while the patient remained immunosuppressed with prednisone and azathioprine. This was accompanied by depressed immunologic tests. Discontinuation of azathioprine and addition of chemotherapy (bleomycin and vincristine), while continuing prednisone to maintain functional survival of renal allograft, has led in this patient to regression of extensive cutaneous and suspected pulmonary Kaposi's sarcoma lesions. The possible importance of a depressed immunosurveillance mechanism and activation of latent oncogenic virus by the presence of an allograft in the de novo appearance of Kaposi's sarcoma in transplant recipients is briefly discussed.


An approach to the serodiagnosis of human lung cancer. Cell culture lines reactive as antigens with tumor patients' sera. A solid-phase radioimmunoassay technique was used to quantitate antigen-antibody reactions between various human cell lines and lung cancer patients' sera. Four human fetal lung cell lines and four human tumor cell lines were more or less reactive as antigens. Failure to obtain exact correspondence between reactions with these cell lines indicates that more than one antigen may be required for detecting specific antibodies to the various lung tumor types. These results suggest that serum antibody detection might be a feasible approach to the immunodiagnosis of lung cancer at stages when the tumor masses are relatively small.


Radioimmunoassay of serum alpha-fetoprotein in patients with different maliganant tumors. The level of serum alpha-fetoprotein (AFP) was estimated by radioimmunoassay in 153 normal healthy Malysians of different ethnic groups. The mean level was 7.5 In1/ml (SD 2.28InU/ml). Among 330 patients with malignant tumors, 11 had increased levels of AFP. The only patient who had hepatoma had a very high level of serum AFP. High levels were also found in three of four patients with dysgerminoma of the ovary, in the only two patients with carcinoma of the testis, and in one patient with secondary carcinoma of the humerus of unknown origin. Lower, but significantly increased levels were observed in one patient (of 48) with breast carcinoma, one patient (of 8) with basal cell carcinoma of the nose, one patient (0f 27) with carcinoma of the lung, and one patient (of 59) with nasopharynegeal carcinoma.


A comparative clinical trial of 5-azacytidine and guanazole in previously treated adults with acute nonlymphocytic leukemia. Adults with previously treated acute nonlymphocytic leukemia received either 5-azacytidine or guanazole in a randomized study. Eighteen patients were treated with 5-azacytidine at a dosage of 200-250 mg/m2/day X 5 intravenously (i.v.) and six achieved a remission (five complete). The median duration of complete remission was 100 days. Among the 12 patients who received guanazole, at a dosage of 25-30 g/m2/day X 5 by continuous i.v. infusion, only one partial remission ensued. Pm 600 WBC/mm3) than nonresponders (median 1700 WBC/mm3). Both the time taken to reach the nadir white blood coung (median, 14 days) and theduration of the nadir (median, 17 days) were long after each course of 5-azacytidine, particularly for those patients who achieved a remission. Principal toxicities seen after 5-azacytidine administration were gastrointestinal tolerance, fever, and neuromuscular toxicity. Fever was the principal toxicity observed after guanazole therapy; one patient developed erythema nodosum with arthralgias and another, recurrent pulmonary infiltrates. Survival from the start of therapy was clearly longer for the patients receiving 5-azacytidine (median 140 days) because of the prolongation of survival seen in the responding patients (median 266 + days). 5-Azacytidine has significant activity as an induction agent in adults with acute nonlymphocytic leukemia, but guanazole does not appear to be of particular value for patients with this disease.


Phase II evaluation of bleomycin. A Southwest oncology Group study. Bleomycin given intravenously (i.v.) or intramuscularly (i.m.) in twice-weekly doses of 10 mg/m2 was evaluated for efficacy and toxicity in 382 patients. Responses were observed in 11/27 Hodgkin's diseases, 10/30 lymphomas, 9/22 squamous cell cancers of ectodermal origin, 12/26 germinal cancers, and 3/8 renal adenocarcinomas. The i.m. route is less likely to casue pulmonary toxicity or hypotension than the i.v. route. Advanced age and total doses exceeding 200 mg were associated with a higher risk of lung toxicity. All responders had shown at least improvement upon receiving 200 mg; higher total doses should be used only in responding patients.


Early myocardial ischaemia: evaluation of the histochemical haematoxylin-basic fuchsin-picric acid (HBFP) staining technique. A series of experiments, carried out to evaluate the histochemical method for the morphological diagnosis of early stages of myocardial ischaemis (HBFP) is reported. The experiments were performed on dog hearts in which ischaemia was induced by coronary artery ligation for different periods of time. The original procedure or modifications of the HBFP-technique, including different staining, rinsing, and differentiation times, the use of different commercial brands of chemicals and preparatory changes of routine histological procedures such as fixation, embedding, cutting, and mounting, did not give satisfactory results. False positive and negative staining was frequent. Very equivocal results were obtained on serial sections of ischaemic tissue samples. Therefore this method was regarded as unreliable and non-reproducible.


Microtubular number in the tractus hypophyseus of newborn normal rats and newborn rats with congenital diabetes insipidus. Evidence has already been adduced suggesting that an increase in microtubular number occurs in the tractus hypophyseus of rats stressed by the administration of hypertonic saline, and of rats with congenital diabetes insipidus (CDI). Since the tractus hypophyseus in these animals shows high secretory activity, it seems likely that the microtubular increase reflects the participation of microtubules in axoplasmic transport. To exclude, however, a congenital microtubular abnormality in CDI, affected newborn rats were examined. In these, the microtubular number was normal, thus suggesting that the increase in microtubular number seen in adult animals was not a congenital morphological abnormality. However, by 4 days of age there was a slight but statistically significant increase in microtubular number in affected rats, a change probably attributable to increased secretory activity.


Effect of Fv-1 gene product on synthesis of N-tropic and B-tropic murine leukemia viral RNA. The production of virus and the synthesis of virus-specific RNA has been studied in Fv-1n/n (NIH/3T3, SIM) and Fv-1b/b (BALB/3T3, SIM-R) cell lines after infection with N- or B-tropic MuLV. It was found that virus production, measured by reverse transcriptase activity in the medium, was 70-100 fold lower in cells resistant at the Fv-1 locus than in permissive cells. The virus-specific RNA, detected by hybridization. In RNA excess with complementary DNA, was reduced by approximately 70-100 fold in cytoplasm of resistant cells compared to permissive cells. A reduction of the same magnitude was observed in the levels of virus-specific RNA extracted from nuclei of resistant cells. Our data therefore show that virus-specific RNA levels are reduced in cells nonpermissive at the Fv-1 locus, suggesting that restriction of the Fv-1 gene product occurs at the level of transcription of the viral genome or at a pre-integration step, or, alternatively, that the RNA transcripts are rapidly degraded after their synthesis.


Colchicine effects on neurosecretory neurons and other hypothalamic and hypophysial cells, with special reference to changes in the cytoplasmic membranes. The morphological effects of colchicine on the entire neurosecretory (NS) tract and on various hypothalamic nuclei have been studied. The perturbations in axonal flow, indicated by the accumulation of NS material, coincide with fragmentation of the cytoplasmic membranes, i.e. the Golgi apparatus and the endoplasmic reticulum, whereas the neurotubules remain relatively well preserved. Autophagic destruction of NS material is observed along the entire length of the NS fibres. The rapid and systematic changes in the axoplasmic reticulum, known to store calcium, lead us to envisage a role for this system-similar to that of the sarcoplasmic reticulum-in controlling the transport of NS vesicles. The junctional zone between the stalk and the neural lobe seems to play a particular role in the transport of NS material to the posthypophysial terminals of the NS axons. Colchicine provokes an increase in dense-cored vesicles in most of the neurons of the other hypothalamic nuclei studied: arcuate, suprachiasmatic, periventricular and ventromedial. Membranous alterations are also observed in these sites. Colchicine administered to animals which were hypothyroid, castrated or adrenalectomized, reveals stimulated neurons, identified by their excessive content of dense-cored vesicles. These neurons display no specific localization, for they occur in all hypothalamic nuclei, irrespective of the stimulation. The frequency of stimulation of neurons of the periventricular nucleus is striking.


Message and non-message sequences adjacent to poly(A) in steady state heterogeneous nuclear RNA of HeLa cells. Highly purified steady state heterogeneous nuclear RNA from HeLa cells has been prepared by a new procedure. Detergent-washed nuclei are disrupted in 0.4 M ammonium sulfate, which also disociated contamination polysomes. The hnRNA remains bound to chromatin, which can be pelleted by gentle centrifugation. Ribonuclease inhibitors permit the preparation of very high molecular weight nuclear RNA. The hnRNA was cleaved with alkali. The poly(A)-containing fragments were separated from those containing oligo(A), and a cDNA copy was prepared. Hybridization of this nuclear cDNA to cytoplasmic mRNA showed that the scarce (complex) message sequences make up a larger proportion of nuclear RNA than of cytoplasmic RNA. In addition, at least 30% of the poly(A)-adjacent sequences in nuclear RNA have no apparent counterparts in the cytoplasm. cDNA prepared from hnRNA sedimenting faster than 45S under denaturing conditions gives similar results, showing the presence of both message and non-message sequences in very large transcripts. cDNA complementary to mRNA was separated into the abundant and scarce sequences, and hybridized separately to the poly(A)-adjacent sequences in nuclear RNA. The hybridization of the abundant sequence cDNA was used to set an upper limit on possible cytoplasmic contamination. Hybridization of the scarce cytoplasmic sequences are represented in nuclear RNA approximately once per cell.


Right ventricular volume characteristics before and after palliative and reparative operation in tetralogy of Fallot. Right heart volume data were obtained in 63 patients with tetralogy of Fallot. The patients were divided into three groups: 1) preoperative tetralogy (N=34); 2) post shunt procedure (N=14); 3A) post repair without outflow patch (N=10); 3B) post repair with an outflow patch (N=8). In Group 1 right ventricular end-diastolic volume (RVEDV), RV ejection fraction (EF), and RV systolic output (SO) were all mildly depressed. In post shunt patients, RVEDV was normal but RVEF remained depressed. RVEDV and RVSO increased following a shunt procedure, and these variables were larger in patients with a large versus a small shunt. In Group 3A RVEDV, RVEF, and RVSO were normal. In contrast in patients in Group 3B, RVEDV was increased averaging 177 +/- 15% of normal RVEF was depressed averaging 0.45 +/- 0.04, and RVSO was normal. RV size and pump function are abnormal in patients whose operation requires an outflow tract patch and the factors which may contribute to these abnormalities include a higher RV peak pressure, pulmonary incompetence, and a larger noncontractile outflow tract. Longitudinal studies relating these variables to clinical performance and exercise testing will be important in assessment of the importance of these abnormalities.


Lymphocyte sensitization to Aspergillus fumigatus antigens in pulmonary diseases in man. In vitro studies of T-lymphocyte responses using five different batches of Aspergillus fumigatus antigens, were undertaken in twelve patients with A. fumigatus related lung disease and in three normal controls. Using a leucocyte migration method, five of the twelve patients showed significant inhibition of leucocyte migration with a migration index of 0-80 or less to A. fumigatus, but in only three was this demonstrated with more than one batch of antigen (one patient with aspergilloma and two with bronchopulmonary aspergillosis). The same antigens were used in lymphocyte transformation tests. Only two patients, one with aspergilloma and one with bronchopulmonary aspergillosis, showed clearly significant transformation although there were several borderline results. Only three patients had evidence of delayed skin responses to A. fumigatus antigens in vivo, one with aspergilloma, one with bronchopulmonary aspergillosis and one with atypical bronchopulmonary aspergillosis. Two of these three patients also had one or more positive in vitro test results. Thus T-lymphocyte sensitization to A. fumigatus as demonstrated by these in vitro methods, although present in occasional patients, was not clearly related to any one particular clinical syndrome in this small group of patients with aspergillus related pulmonary disease. There was, however, one of the three aspergilloma patients with positive lymphocyte transformation to all five batches of antigens and having higher transformation indices than in any other patient. This suggest that lymphocyte studies should be extended in this group. In contrast to the frequent negative results using A. fumigatus antigens, evidence of T-lymphocyte sensitization to either Candida albicans or Mycobacterium tuberculosis or both, was shown by positive delayed hypersensitivity skin responses and in vitro inhibition of leucocyte migration in the majority of the patients, despite which lymphocyte transformation was often negative. The possibility of impaired capacity to transform is supported by the finding of an impaired response to phytohaemagglutinin (PHA) in four of the twelve cases. This information, together with other data discussed, helps to complete the contrasting immunological profiles seen in different Aspergillus fumigatus related lung diseases.


Comparison of the allergenic properties of bee venom and whole bee body extract. The allergenic properties of bee venom and whole bee body extract were compared by in vivo and in vitro tests. The majority of patients with known bee sting sensitivity had positive intracutaneous skin test reactions with bee venom and had bee venom specific IgE in their sera. Of seventeen patients with ppsitive bee venom skin tests, nine had positive tests with whole bee body extract. Of thirty sera containing elevated levels of bee venom specific IgE obtained from untreated patients, fourteen sera contained whole body specific IgE but in much lower titres. In RAST inhibition experiments using both bee venom and whole bee body extract as coupling antigens, bee venom was a more potent inhibiting antigen than whole body extract. From these experiments we conclude that bee venom is a more potent allergen than whole bee body extract.


Purification of alpha1-fetoprotein.

Purification of alpha1-fetoprotein. Previously published methods for purifying alpha1-fetoprotein are inadequate because they either do not yield a completely pure product or they cause some denaturation. We present a method that does not have these serious disadvantages, and with which alpha1-fetoprotein was purified by sequential use of concanvalin A affinity-chromatography, preparative gel-electrophoresis, and immunoabsorption with anti-albumin antibody covalently coupled to Sepharose 4B. The purity of the product was monitored by discontinuous polyacrylamide-gel electrophoresis and counterimmunoelectrophoresis, both of which must be used to ascertain what proteins are present at each step of the purification.


Binding of 3,5-diiodotyrosine to serum proteins. The reversible binding of 3,5-diiodotyrosine (DIT) to human and bovine serum protein and to purified human serum prealbumin and human and bovine albumin has been studied by equilibrium dialysis. Maximum binding occurred at pH 8.6-9.0. Human serum bound DIT less than did bovine serum. Adult ox and fetal calf sera showed similar binding. The main DIT-binding protein of human serum was prealbumin. It showed a single affinity site with a Ka of 0.85 X 10(6) M-1 at pH 8.6 and 0.40 X 10(6) M-1 at pH 7.4. The affinity constant of serum albumin for DIT was 2.8 X 10(3) M-1 at pH 8.6. The elevated binding of DIT to bovine serum is essentially due to albumin whose affinity constant for DIT is 16-times higher than that of human serum albumin. Fetuin was not responsible for any noticeable DIT binding in fetal calf serum.


A solid-phase radioimmunoassay for human beta2-microglobulin. A simple and relatively rapid as well as highly reproducible determination of beta2-microglobulin is described for general laboratory use. The method is based on a solid-phase radioimmunoassay paper discs as a solid-phase material. The assayable range is approximately in the range of 2 ng to 20 mug of beta2-microglobulin with a precision of approx. 5 percent. This assay requiring only 0.05 ml of urine can be completed within 20 h. Urine in cadmium-exposed workers and residents in cadmium-polluted districts including patients with itai-itai disease were examined and considerable amounts of beta2-microglobulin were observed in these urines.


Liver reaction to tissue injury: do therapeutic measures trigger the acute phase reaction? In 30 healthy people we investigated the hypothesis that therapeutic measures trigger the acute phase reaction. In order of severity we analyzed: (1) the effects of an intramuscular injection of 2.5 ml 0.14 molar NaCl (6 patients), (2) the effects of an indwelling venous catheter for 24 h (7 patients), and for 72 h (8 patients), (3) the effects of cardiac catheterization (arterial and venous catheterization) carried out in 9 patients. In clinical investigations of the acute phase reaction, an intramuscular injection and a 24- or 72-h indwelling catheter, brought in by venipuncture, are acceptable because they do not provoke an acute phase reaction. A venous cutdown associated with an arterial puncture is not acceptable because this injury evokes a positive acute phase reaction. This was also observed in two patients who developed phlebitis during a 24-h indwelling venous catheter. Only a positive acute phase reaction was found, not a negative one.


Aspects of the secondary antibody response to ox insulin in the Hartley guinea-pig; the use of chemically modified ox insulin to delineate the antigenic determinants of ox insulin. The use of a quinea-pig model to study the immunogenicity of the insulin molecule is presented. The Hartley guinea-pig has been shown consistently to form antibody to ox insulin, when given in a water-in-oil emulsion containing pertussis vaccine as adjuvant. After log transformation of standardized antibody titres to iodo-ox insulin, a valid statistical comparison of the antibody response to different ox insulin preparations could be made. Antibody cross-reacting with ox insulin, but not iodo-ox insulin, was also detected. The quantity of one type of antibody was complementary to the other, an observation compatible with determinant competition having occurred during the immune response. From the results of cross-reactivity experiments using N-triacylated ox insulins and human insulin, it was shown that antibody cross-reacting with iodo-ox insulin had most probably been produced to a localized area of the molecule.


A direct macrophage migration inhibition test applied in man. A direct macrophage migration test is described, which proved to be useful for the detection of cellular hypersensitivity in man. We used a modification of the method described by Hughes & Paty (1972). In this method the MLR is abolished by 100 rad gamma-irradiation of the peritoneal exudate cells prior to pooling with human lymphocytes. Experiments with various intensity of irradiation, PPD, muscle antigen and encephalitogenic factor were performed to check this method. In a pilot study lymphocytes of patients with diseases of the central or peripheral nervous system or of muscle were tested. This proved that a hypersensitivity of Ef was present in various diseases of the CNS, while in muscle diseases positive tests were found using muscle antigen.


Membrane characteristics of established human T- and B-cell lines. Cross-reactivity of human T-antigenic determinants with peripheral lymphocytes of non-human primates and the presence of MLC antigens on cultured T-cell lines. Human T-cell lines (MOLT-4 and SOMMER-T) were injected into rabbits and monkeys (stumptail and squirrel monkeys). Rabbit anti-MOLT serum was absorbed with human liver and cultured B cells. Absorbed anti-MOLT serum was cytotoxic to lymphocytes of baboons and stumptail monkeys. Rabbit anti-MOLT and SOMMER-T sera after absorption with liver and B cell showed florescent ring formation in baboon and stumptail lymphocytes by using immunoflurescence techniques. On the other hand, antisera against MOLT and SOMMER-T  cells in stumptail and squirrel monkeys were not only cytotoxic to MOLT and SOMMER-T cells, but also to other T-cell lines, CCRF-CEM and CCRF-HSB-2 cells. Cultured B-cell lines stimulated allogeneic and xenogeneic (rabbit and monkeys) lymphocytes far better than cultured T-cells lines did. T-cell lines, CCRF-HSB-2 and SOMMER-T (number 8402), gave small but significant stimulation to allogenic lymphocytes especially in the presence of foetal calf serum. MOLT-4B failed to stimulatie allogeneic lymphocytes. When lymphocytes of non-human primates and rabbit were cultured with human B- and T-cell lines in the presence of foetal calf serum, CCRF-HSB-2 and SOMMER-T cells stimulated xenogeneic lymphocytes of rabbit, squirrel monkey and stumptail monkey. MOLT-4 cells stimulated lymphocytes of baboon to some extent. These results indicate that although cultured B-cell lines had more mixed lyphocyte reaction (MLR) stimulating structure than T-cell lines, the latter still maintained some stimulating structure on the membrances.


Effect of vagotomy on anaphylaxis in the rat. Vagatomy or atropinization protects rats from lethal anaphylaxis and histamine shock. These procedures do not significantly affect production of anaphylactic antibodies or release of histamine following induction of anaphylaxis. The response of the intestinal microvasculature during anaphylaxis is observed to consist of an early brief increase permeability followed by prolonged and irreversible damage. Cholinergic blockade abolishes the latter and markedly reduces deposition of colloidal carbon and immune complexes in the vessel. The possible relevance of these observations to development of chronic immunological vascular disease is discussed.


Determinant competition during the immune response to N-acyl derivatives of ox insulin in the Hartley guinea-pig. Subgroups of female Hartley guinea-pigs were immunized with N-carbamylated ox insulin, N-maleylated ox insulin, N-phthaloylated ox insulin, or with the crystalline ox insulin from which the N-acylated insulins had been prepared. The immunogens were administered in water-in-oil emulsions containing pertussis vaccine as adjuvant. Sera obtained 20 days after secondary immunization were assayed for their antibody titres to iodo-ox insulin and their insulin-binding capacities. The data were log transformed for statistical comparison. N-carbamylated ox insulin seemed to be as immunogenic as crystalline ox insulin and no specific carbamyl hapten antibody could be found. N-maleylated and N-phthaloylated ox insulins yelded significantly less antibody cross-reactingwith iodo-ox insulin, but produced a complementary quantity of specific maleyl and phthaloyl hapten antibody respectively. Thus it was shown that in the system used the immune response was partitioned between different determinants, ox insulin and its N-acylated derivatives being equipotent immunogens.


The disappearance of enzyme-inhibitor complexes from the circulation of man. 1. Complexes of human trypsin and human granulocyte elastase with alpha1-anti-trypsin and alpha2-macroglobulin were isolated and injected intravenously into human volunteers. 2. The elimination of alpha2-macroglobulin complexes with trypsin and elastase followed single-exponential functions with half-lives of 9 and 12 min respectively. The complexes showed no tendency to dissociate. 3. Complexes of alpha1-anti-trypsin with trypsin persisted in the circulation much longer, with a half-life of 3-5 h; complexes of alpha1-anti-trypsin with elastase had an intermediate half-life of 1 h. 4. Dissociation was observed of alpha1-anti-trypsin complexes with transfer of trypsin and elastase to alpha2-macroglobulin. 5. Dialysable radioactivity appeared in the urine soon after the injection of alpha2-macroglobulin complexes, which suggested a breakdown of complexes by cells in the reticuloendothelial system. Radioactivity over the liver achieved maximum values within 30-40 min after the injection of alpha2-macroglobulin complexes but not until 50-70 min after the injection of alpha1-anti-trypsin comlexes. 6. These results support the concept of a key position for alpha2-macroglobulin in the protective mechanisms against endogenous proteases.


A guide to the synaptic analysis of the neuropil. A morphological analysis of the organization of the gray matter in the central nervous system depends on the discovery of consistent repetitive patterns. Without these, the gray matter remains a chaotic jungle. An hypothesis derived from the study of a few simple regions has been developed to serve as a guide in finding these patterns. It states that all nerve fibers and terminals arising from a particular group of nerve cells, or, more precisely, a particular nerve cell type, display similar axoplasmic configurations despite variations in size and shape of the terminations. This hypothesis is reminiscent of the so-called Dale's principle that a nerve cell makes use of the same transmitter at all of its branches or terminations. These apparent rules of uniformity or congruity merely reflect the functional integrity of the nerve cell and the role of its parts in the nervous system. But as an hypothesis, it needs to be tested, and it needs to be tested anew in each region, since exceptions to the assumed rule can be expected. It is therefore proposed as the first working hypothesis in each new region. If it should prove to be true in general, it will facilitate and rationalize the analysis of the gray matter, as it has already done in the cerebellar cortex and the deep cerebellar nuclei. If it should prove to be false in a few regions, the analysis will become more difficult, and additional modes of marking nerve endings will have to be used. Experimental methods for identifying nerve terminals can be translated from the light microscopic to the electron microscopic level, but there are significant drawbacks at both levels: lack of precision, destruction of fibers of passage, and rapid evolution of the degenerative process may greatly restrict their usefulness. Labeling with tritiated amino acids or transmitters, or with horseradish peroxidase, provide new methods for tracing interneuronal connections at the electron microscopic level. These have the advantages of high specificity, nondestructiveness and a physiological mode of selective marking. However, they do not offer a solution to the problem of short-range connections. For these, careful reconstructions of serial sections may prove necessary, as Sjöstrand (1974) has demonstrated in a remarkable paper on the retina. The aim of all these methods is to discover patterns of synaptic connectivity in order to map the cellular organization of the nervous system. In the foregoing, nothing was said about synapses other than those articulating axons with somata or dendrites and their appendages. Clearly the same principles of recognition apply to axo-axonal and dendro-dendritic synapses. Although the synapses that have been considered here are chemical synapses, the same questions regarding the identity of the partners in electrotonic junctions must be asked as well.


Axonal transport of vesicles carrying (3H)serotonin in the metacerebral neuron of Aplysia californica. The axon of each unipolar metacerebral neuron bifurcates into two large branches within the cerebral ganglion quite close to the cell body, and each branch exists from the ganglion in a separate nerve, either the lip nerve or the cerebrobuccal connective. Because the serotonergic vesicle appears to be more stable to isolation than the cholinergic vesicle and because serotonin, like other transmitter molecules with primary amino groups, can be fixed into tissue with aldehyde fixatives, we were able to show that serotonergic vesicles, originating from the cell body, are transported in both branches of the axon by fast axonal transport. We also described some experiments suggesting that the membranes of the serotonergic vesicles being transported were newly synthesized in the cell body and contained vesicle-specific glycoproteins. This is important because it permits us to estimate the degree of the neurons' biosynthetic activity involved in the formation of these vesicles. Finally, we took advantage of the axon geometry of the metacerebral cell to perturb the dynamics of the transport system. Cutting one branch abruptly increases the number of vesicles traveling along the uncut axon. Using this manipulation, we found that when twice the amount was being transported in the uncut nerve, [3H]serotonin was distributed more distally, suggesting that on the average, vesicles were displaced twice as far per unit time. We therefore proposed a mechanism for axonal transport which involves the binding of vesicles to stable tracks as a rate-limiting step. When attached, vesicles move at a constant, fast rate.


[Post-MOPP chemotherapy of Hodgkin's disease (author's transl)]. In 18 patients with advanced Hodgkin's disease, refractory to previous 'C-MOPP' treatment, a new therapeutic protocol of adriamycin, DTIC, CCNU and bleomycin was introduced. Three patients who had previously failed to respond to any other chemotherapy failed also to respond to the new one. But two complete and 11 partial remissions were obtained in the other 15 patients who had previously responded to C-MOPP before becoming refractory to it. Two patients had further progression of the disease during the new treatment.


[Demonstration of mycoplasma by direct fluorochromation with acridine derivatives (author's transl)]. Vital fluorochromation of mycoplasmas can be obtained with acridine derivatives. Stained with acridine-orange und 9-aminoacridine chloride-1-hydrate mycoplasmas appear as yellow (green) fluorescing homogeneous spots or discs under the fluorescence microscope, the exact appearance depending on the concentration of the fluorochrome and the absorption by nucleic acids. Non-specific supplements and undermixed bacteria have significantly different fluorescence. Depending on the metachromasia of acridine-orange and on double-fluorescing with acridine-orange and 9-aminoacridine chloride-1-hydrate, the bacterial cell wall is green, the protoplasma red. The method is suitable as a screening test for mycoplasma.


[Comparative studies on the teaching of oral pathology]. This is a self-assessment of teaching effectiveness for teacher, students and teaching-material as well. A curriculum of oral patholology was created and prepared as syllabus, lectures and audiovisual aids. In order to estimate and compare the eductional achievements of different systems, a series of written multiple choice questions were prepared. Only items with an index of discrimination of more than 0,25 were used. The average index was 0,5. The results of a short-term experimetn (1 hour) shows that a teaching sequence in the conventional lecture style was most effective in transmitting knowledge (72% of maximum), audiovisual aids rating second (63%), followed by individual homework study using printed material (48%). The differences were statistically significant in most experiments. In the second type of experiment (long-term-experiment) we registered the time the students spent with lectures over a period of six months. When permitted to chose between the different systems the students preferred the audiovisual method to the lectures. Individual testing of these students shows a positive correlation between knowledge acquired and time spent with the audiovisual machines.


Serum alpha1-fetoprotein in cystic fibrosis. In 38 children and adolescents with cystic fibrosis, in 19 normal children and 62 healthy adults the serum alpha1-fetoprotein concentrations were measured by radioimmunoassay. In cystic fibrosis patients 97.5% and in normal children 95% of the alpha1-fetoprotein values were within the normal range for healthy adults (1--9 ng/ml). Critical judgement of the reported findings in literature and our own results demonstrate that the investigation of alpha1-fetoprotein in the serum cannot serve for detecting homozygotes of cystic fibrosis genes or heterozygote carriers.


Cystic fibrosis: normal volumes of red cells. Model experiments on the pathomechanism of the disease. Normal values of mean cell volume (M.C.V.) and of distribution of single cell volumes (S.C.V.) have been observed in erythrocytes of patients with cystic fibrosis using an electronical particle-volume analyzer (sheath flow detector). From these results a strong defect in red cell salt transport seems improbable. Valinomucin induces shrinking (by increase of K+ permeability) and gramicidin D induces swelling (by increase of Na+ permeability) of normal erythrocytes. Addition of C.F. sweat to normal erythrocytes induces no volume change. From this result no influence of the "C.F. factor" on passive ion permeability is concluded.


Immunological investigations of antigenicity and specificity of soluble collagen fractions. IV. Anaphylaxis and allergy experiments. Collagen prepared from rat, calf, or pig skin was tested on guinea pigs for active cutaneous anaphylaxis. Only one fraction of calf skin collagen gave a slight, positive reaction. No positive reactions were observed in 21 subjects submitted to patch tests and 12 subjects submitted to scratch tests with pig skin collagen. Therefore preparations of pig skin collagen may be used for wound treatment without any risk.


Evaluation and management of the child with delayed pubertal development. The physical and hormonal changes of puberty are presented and the wide range of ages at which the pubertal process may begin is emphasized. The great variability in the timing of onset of adolescence, its rate of progression, and the age of completion are detailed. The causes of delayed adolescence in males and females are considered. The most common form of delayed adolescent development is termed constitutional delay in growth and development, which may occur sporadically, or may be the familial pattern of growth and development or may reflect a suboptimal nutritional environment. The evaluation of such children, including appropriate historical review, physical examination, and laboratory assessment, is outlined. In most patients with constitutional delay in growth and development, strong reassurance is sufficient therapy. In other subjects, treatment with androgens (boys) or estrogens (girls) may be indicated. In patients with primary systemic diseases accociated with delayed maturation, specific treatment which eradicates the illness will often be followed by resumption of growth and development. In subjects with primary disorders of the hypothalamus, pituitary, or gonads, replacement therapy with androgens or estrogens is indicated. If gonadal function is intact, these patients may eventually become fertile with appropriate use of hypothalamic and/or pituitary hormones.


Studies of the "e" antigen in acute and chronic hepatitis. Sera from well individuals, including controls and asymptomatic HBsAg carriers, post-transfusion hepatitis cases, and chronic active liver disease patients were examined for the presence of "e" antigen and e antibody by rheophoresis. Our data confirm the specific association between the e determinant and hepatitis B infections and indicate that e antigen is closely associated with evidence of chronic hepatic dysfunction, in contrast to the association of e antibody with hepatic normalcy in HBsAg carriers. However, these correlations are not absolute and, therefore, it should not be inferred that all e antigen-positive individuals will develop chronic hepatitis nor, conversely, that presence of e antibody invariably protects against the development of chronic hepatitis.


Differential distribution of hepatitis B surface antigen and hepatitis B core antigen in the liver of hepatitis B patients. One hundred liver biopsies from 100 patients with clinical presumptive diagnosis of hepatitis were examined by immunofluorescence for the presence of hepatitis B surface antigen (HBSAg) and hepatitis B core antigen (HBcAg). Of the 60 HBsAg-positive livers, 51 were diagnosed as chronic hepatitis on histological grounds, 6 as acute hepatitis, and 3 as "near-normal liver." From the 60 tissue-positive cases, 3 subjects were HBsAg seronegative. HBcAg was detected in 44 livers, all of which also had HBcAg in the localized in the cytoplasm and the membranes of the hepatocytes, and HBcAg in the nuclei and in 4 cases also in the cytoplasm. Predominant HBsAg expression in the cytoplasm was observed in near-normal liver, chronic persistent hepatitis, and cirrhosis with little activity. This correlated with the amount of ground glass hepatocytes in the biopsies. HBcAg and membrane-localized HBsAg were minimal in those conditions. HBcAg was most prevalent in patients with chronic aggressive hepatitis and active cirrhosis treated with immunosuppressive drugs, whereas the amounts of HBsAg and HBcAg in nontreated patients of those two groups and in acute hepatitis with signs of transition to chronicity were almost equal. HBsAg expression in liver cell membranes was most prominent in active forms of chronic hepatitis (chronic aggressive hepatitis and in active cirrhosis) and in acute hepatitis with signs of transition to chronicity. This observation correlated in the presence of HBcAg in the biopsies of those patients. In acute hepatitis both HBsAg and HBcAg were detected rarely and no membrane expression of HBsAg was observed. The over-all results show a significant relationship between the different degrees of accumulation of HBsAg and HBcAg in the liver and the various histological types of hepatitis and further suggest an interplay of both hepatitis B virus and host immune response in the development and pathogenesis of hepatitis B.


Occurrence of alpha-fetoprotein, Regan isoenzyme, and variant alkaline phosphatase in the serum of a patient with gastric cancer. Regan isoenzyme, variant alkaline phosphatase, and alpha-fetoprotein were found in the serum of a patient with gastric cancer. The histology of the tumor was tubular adenocarcinoma. There were metastases in the retroperitoneal lymph nodes, but not in the liver. The liver was normal microscopically, with no evidence of bile duct obstruction. alpha-Fetoprotein in the tumor tissue was detected by immunoprecipitation reaction in agar. Regan isoenzyme and variant alkaline phosphatase were also detected in the tumor tissue and total alkaline phosphatase activity of the tissue was very high. These findings suggested their tumor origin.


Direct method for estimating the total gastrin cell number in the stomach of rats. An original and simple method for counting the total G cell number in the stomach of small rodents is described. Staining the G cells with the peroxidase-antibody technique and counterstaining the nucleus make it possible to correct the nuclear overestimation. Counting directly the total number of G cells in sections from the entire stomach avoids errors due to tissue retraction and to unprecise delineation of the antro-fundic limit.


A "new" blood factor, C1, demonstrated with extracts of seeds of the Korean Clerodendron trichotomum Thunberg. By absorbing extracts of seeds of the Korean Clerodendron trichotomum Thunberg with selected human group O red cells, a lectin has been prepared, which defines a hitherto undescribed specificity, designated C1, defining individual differences in human red cells. The specificity C1 appears to characterize a structure associated with the A-B-H-Le marcomolecule, both of red cells and of saliva, which is distinct from the combining groups for A,B,H and Le. Moreover, the reactivity of red cells with anti-C1 lectin is destroyed by treatment of the red cells with proteolytic enzymes, unlike the reactions for A,B,H and Le.