Relationship of cellular potential hemolysin in group A streptococci to extracellular streptolysin S. The relationship of streptolysin S (SLS) and a cellular potential hemolysin (PH) activatable by sonication was examined in strain C203S (a known high producer of SLS), its SLS(-) mutant (C203U), and in 20 other group A streptococci of various M and T serotypes. All strains shown to form SLS (ribobycleic acid (RNA)-core SLS) contained PH. The two strains lacking PH were the only ones that did not produce SLS In strain C203S, formation of SLS bycells incubated with RNA-core for 60 min at 47 C in a nongrowth basal medium (Bernheimer's basal medium) was followed by a marked decrease (99.6% loss) of PH titer. Without stimulation of SLS production by addition of RNA-sore, the same incubation resulted in a progressive but less marked fall (38.8%loss in 60 min) of PH titer: these cells produced disproportionately low titers of SLS on subsequent addition of RNA-core. This effect of prior incubation in Bernheimer's basal medium on SLS titer was partially nullified by use of fresh medium after 30 min, but not after 60 min, and did not occur during 60 min of incubation at OC. These results provide additional evidence for a precursor-product relationship between PH and SLS. They also suggest that a medium factor (or factors) is utilized or destroyed at 37 C and that this factor is essential to both the stability of PH and its efficient conversion to SLS.


Erythrocyte antinuclear antibodies in sera of chickens hyperimmunized with group A streptococcal vaccine. Chickens hyperimmunized with group A streptococcal vaccine often synthesize high levels of antibody to group A streptococcal carbohydrate (SACHO). Indirect immunofluorescent analysis revealed that sera of these chickens also contain antinuclear antibodies capable of reacting with chicken erythrocyte nuclei (EANA) at titers up to 2,560. Removal of the anti-SACHO antibodies from hyperimmune serum did not significantly reduce EANA titers, indicating that anti-SACHO antibodies are not responsible for the EANA reactions. The time course for antibody development as well as the immunoglobulin class distribution of EANA and anti-SACHO antibodies were very similar. Localization of EANA activity to Fab fragments indicated that the immunfluorescent reaction represents an antigen-antibody reaction. Findings point out an important consequence of using chicken immunoglobulin Y in immunofluorescent assays. The purified Fc fragment of immunoglobulin Y is cytophilic for the substrate when tested at high concentrations, which suggests that some cytophilic immunoglobulins are involved in the reaction.


Antigenicity of converting phages obtained from Clostridium botulinum types C and D. Phage conversion of toxigenicity in Clostridium botulinum types C and D was accomplished by using nontoxigenic strains and phages purified from plaques. Although the morphology of the converting phages seemed to be the same, they were divided into three groups on the basis of their conversion spectrum. The first group consists of phages obtained from toxogenic strains C-Stockholm and C-468. The second group consists of phages from strains D-1873 and C-203. The third group consists of phages from strains D-South African and D-4947. These converting phages were also classified into the same three groups by a neutralization test with specific antiphage sera. Cross-neutralization, however, was observed between phages belonging to group 1 and group 2,by both the neutralization test of converting ability and by a plaque experiment in which the surviving rates of phages were calculated after treatment with each antiphage serum. The antigenic differences among these converting phages should probably comprise one of the reasons for the existence of the specific infection spectrum in C. botulinum types C and D.


Spontaneous interaction in vitro between lymphocytes and syngeneic peritoneal macrophages of mice. Ficoll-purified lymphocytes (peritoneal, splenic, or thymic) and macrophages (peritoneal) from Toxoplasma-immune and normal female NMRI mice were used. Suspensions of washed cells were made in medium 199 containing 20% heat-inactivated normal calf serum. Sixty minutes after the adherence of 10(5) macrophages to cover slips in Leighton tubes, lymphocytes were added in various concentrations. The mixed cellular population was then incubated at 37 C. Eighteen hours later, most of the lymphocytes were firmly attached to macrophages to form rosettes. This cellular interaction, which was temperature, cell ratio, and time dependent, occurred in the absence of any particular antigenic stimulation. Morever, the reaction was cytotoxic only for adhered lymphocytes as judged by staining with 0.2% trypan blue. Splenic and thymic lymphocytes were bound in significantly greater number than peritoneal lymphocytes. Incubation of macrophages for more than 48 h at 37 C before the addition of fresh lymphocytes markedly reduced rosette formation. Treatment of macrophages and lymphocytes with mouse anti-immunoglobulin did not affect the reaction. The labeling of lymphocytes with fluorescent anti-mouse sera and the use of nude NMRI mice showed that both B and T cells can form spontaneous rosettes with syngeneic peritoneal macrophages.


T antigen of Streptococcus pyogenes: isolation and purification. A T antigen preparation free of trypsin was obtained by application of CNBr-Sepharose linked to pure trypsin. Purification on an ion exchange chromatography column results in an electrophoretically homogeneous preparation of T protein.


Experimental group B streptococcal infections in mice: hematogenous virulence and mucosal colonization. A group B streptococcus recovered from a blood specimen from a neonate with sepsis was used to evaluate the use of mice for studies characterizing the hematogenous virulence and the asymptomatic mucosal colonization of the vagina or of the respiratory tract by these bacteria. When injected intravenously, the 50% lethal dose for mice was 10(6); however, as few as 10(2) organisms produced septic deaths. In mice undergoing water diuresis, bacteriuria and pyelonephritis were not produced after direct bladder inoculation of the streptococci. Asymptomatic vaginal colonizations that persisted for 12 days were produced in both pregnant and virgin mice. Vaginal colonization before delivery did not result in transmission of infection to litters or in protection against subsequent oropharyngeal colonization in the suckling mice. In mice born of nonexposed mothers, oropharyngeal colonization was produced in both suckling and 3-week-old weaned mice. Whereas infection persisted for 14 days in all suckling mice, clearance occurred in over 50% of the weaned mice by day 14. The use of mice for studies on the virulence of the group B streptococci as well as for studies on the pathogenesis of disease by virulent strains is discussed.


Detection of the K99 antigen by means of agglutination and immunoelectrophoresis in Escherichia coli isolates from calves and its correlation with entertoxigenicity. The common antigen of calf enterotoxigenic strains of Escherichia coli, recently established as the K99 antigen, was studied by means of the slide agglutination test and immunoelectrophoresis. Specific antisera were obtained by absorption of crude antisera with ultrasonicates of autologous cells grown at 18C or by injection into rabbits of the purifies K99 antigen obtained by preparative electrophoresis. The K99 antigen was usually undetectable in calf enterotoxigenic E. coli cultures with capsular K antigens of the A variety grown at 37C on commercially available nutrient agar plates designed for the isolation of Enterobacteriaceae, but was rapidly detectable when grown on a buffered semi-synthetic medium at pH 7.5 (Minca medium). An alternative procedure for the isolation and identification of calf entertosigenic E. coli strains from feces, using the Minca medium, is proposed. K99 was found in 70 of 74 strains of E. coli, the enterotixigenicity of which was established in the ligated gut test in calves. None of the 20 cultures negative in the ligated gut test possessed K99 antigen. The K99 antigen is therefore probably a useful diagnostic tool for the identification of calf enterotixigenic E. coli strains, taking into account that K99 and enterotoxigenicity are controlled by different plasmids.


Immunity to syphilis: passive transfer in rabbits using serial doses of immune serum. The influence of prolonged treatment with serial doses of immune serum initiated either 2 or 48 h before intracutaneous challenge with Treponema pallidum on the subsequent appearance and development of lesions was studied. Whereas treatment with large doses of immune serum initiated 48 h before treponemal challenge caused a delay of 9 to 20 days in the onset of lesions and marked suppression of lesion development. Treatment initiated 2 h before treponemal challenge failed to delay the onset of lesions and only moderately suppressed their development. Lesion development was suppressed as long as treatment was continued. The significance of the results is discussed.


Effects of a streptococcal lipoteichoic acid on host responses in mice. A teichoic acid (TA) extracted from Streptococcus pyogenes 1-RP41 was previously shown to be an immunosuppressant under certain conditions (Miller and Jackson, 1973). The TA has now been shown to be a lipoteichoic acid composed of 40% glycerol, 20% alanine, 13% phosphorus, and 8% glucose, with a variable content of fatty acids. Teh presence of the polyglycerol phosphate backbone and fatty acid was required for maximum immunosuppression of the primary immunoglobulin M response to sheep cells. A complex, nonlinear, time-dependent dosage relationship in suppression of the anti-sheep erythrocyte response in mice was observed. TA depressed the anamnestic response to sheep cells in the mouse and could affect this response whether administered before the primary antigen challenge or immediately before the secondary challenge. In distinct contrast, TA enhanced antibody production to Escherichia coli O55:B5 lipopolysaccharide when assessed by counting plaque-forming cells or measuring antilipopolysaccharide serum titers. The TA failed to stimulate a large uptake of [3H]TdR by murine spleen cells; however, it significantly enhanced the clearance of carbon by the reticuloendothelial system.


Autoimmunity induced in rabbits by rinderpest virus. During rinderpest virus infection in rabbits, 19S cold hemagglutinating antibody against rabbit erythrocytes and 7S anti-nuclear antibody, which reacts with the nuclei and/or the nuclear membranes in immunofluorescent staining, were demonstrated. Virus infection that affected the thymus-dependent immune functions was speculated to act as a trigger for the production of these two autoantibodies.


Degradation of 14C-labeled streptococcal cell walls by egg white lysozyme and lysosomal enzymes. The resistance of native and trypsin-treated [14C] glucose-labeled cell walls to degradation by lysozyme and human lysosomal enzymes was confirmed. In contrast, chemically N-acetylated cell walls undergo significant degradation by these enzymes in the pH range of 4.5 to 5.5 without prior removal of the group-specific carbohydrate. N-acetylation after removal of the group A carbohydrate by formamide extraction renders the cell walls considerably more susceptible to these enzymes than by formamaide extraction alone. It appears, therefore, that unless N-acetylation can occur in vivo, streptococcal cell walls are minimally degraded, if at all, by human peripheral blood leukocytes or lysozyme. Examination of leukocyte extracts from normal subjects and patients with post-streptococcal syndromes revealed no qualitative differences in ability to dissolve streptococcal cell walls.


Nocardia asteroides and Nocardia brasiliensis infections in mice. A model for Nocardia asteroides and Nocardia brasiliensis infections in Swiss white mice has been established without the addition to the inocula of any form of adjuvant. Serial histopathological studies revealed that these two actinomycetes cause lesions that are quite different in their features. An acute suppurative abscess characterizes the lesions of N. asteroides. In the case of N. brasiliensis infections a granuloma is produced in which a striking feature is the presence of large numbers of foam-laden macrophages, although occasional exceptions to this pattern were noted. Electron microscopic studies demonstrated that these macrophages contain within their cytoplasm organisms in varying stages of degeneration. Repeated mortality studies in mice failed to demonstrate differences in mortality rates produced by N. asteroides and N. brasiliensis. Thus, despite relatively trivial biochemical and antigenic differences between these two species of Nocardia, the local pathogenic response is quite different. The presence in the "brasiliensis lesion" of foamy macrophages with intracellular organisms is reminiscent of the histopathological features of lepromatous leprosy and of disseminated Myocobacterium bovis infection when this occurs in the immune suppressed situation. It is possible that N. brasiliensis infection produces a depression of cellular immunity that modifies the local host response to the organism.


Salmonellosis in mice: immunization experiments with salmonella-escherichia coli hybrids. Mice vaccinated with Escherichia coli hybrids experesing Salmonella O antigens were protected against lethal challenge doses of Salmonella strains. Counts of challenge bacteria in mouse organs revealed that a live hybrid vaccine was slightly more efficient than an acetone-killed vaccine.


Mitogenic factor as an in vitro correlate of delayed hypersensitivity in the guinea pig. The relationship of mitogenic factor (MF) to delayed hypersensitivity was examined in the guinea pig. MF was readily produced in vitro by blood leucocytes from animals sensitized with Keyhole limpet haemocyanin in Freund's complete adjuvant. Such animals exhibited delayed skin reactions and macrophage migration inhibition. Treatment with cyclophosphamide markedly reduced serum antibody but caused no diminution of MF or delayed skin reactions. A migration inhibition response was observed 10, but not 20, days after sensitization by means of incomplete adjuvant.


Production of migration inhibitory factor and blast cell transformation by cord blood lymphocytes. Cord blood lymphocytes from 29 normal infants of 28-41 weeks gestational age were tested for blast cell transformation and migration inhibitory factor (MIF) production. Cultures were stimulated with phytohemagglutinin (PHA) and purified protein derivative of tuberculin (PPD) for transformation; with purified PHA, soluble and insoluble concanavalin A as well as PPD for MIF production. 3H-thymidine incorporation by cord blood and by adult lymphocytes were similar in magnitude after PHA stimulation, but differed significantly after PPD stimulation, with negligible increased incorporation by the former and marked increased incorporation by the adult lymphocytes. With the exception of PPD, the stimulating agents used led to MIF production by cord blood lymphocytes. No dissociation was observed between proliferative response and MIF production. Both functions seem to be fully developed at birth.


Subclass typing of IgG antibodies formed by grass pollen-allergic patients during immunotherapy. The subclasses of IgG antibodies formed by grass pollen-allergic patients during immunotherapy were investigated by using a radioallergosorbent test (RAST) and a quantitative immunofluorescence method known as the defined antigen substrate spheres (DASS) system. By the use of rabbit antisera directed against the subclasses of IgG, the specificity of which was checked in the passive hemagglutination and immunofluorescence techniques, it was shown that a relatively high proportion of the grass pollen-specific antibodies belonged to the IgG4 subclass. Apart from the high binding activity of IgG4 which increased during treatment, a moderate binding activity of the other subclasses was also found. Binding of all subclasses increased slightly in the pollen season and could be specifically blocked by perincubation with soluble grass pollen extract. The results of the IgG4 binding, determined in vitro with the DASS system, and the blocking activity of the sera, determined in vivo by skin tests are suggestive for a relation between these activities. Also in the group of patients with a low IgE-RAST score, the skin reactivity decreased as the IgG4 binding activity increased.


Detection of IgG antibodies against pigeon intestinal mucosa antigens in pigeon breeders' sera using the immunofluorescent technique. The serum of 102 pigeon breeders was examined for circulating IgG antibodies against antigens localized in pigeon intestine by the indirect immunofluorescent technique. 15 symptomatic breeders with a positive response to inhalation challenge had antibodies against mucosa tissue. The majority of asymptomatic breeders demonstrated either a fluorescence of intestinal excrements combined with a weak fluorescence of mucosa tissue or had no detectable antibodies in their serum. By the indirect immunofluorescence, antibodies were found more frequently than by immunoprecipitin reactions in agar.


Nonspecific immunosuppression induced in mice with killed Pasteurella haemolytica in Freund's complete adjuvant. Killed Pasteurella haemolytica type I in combination with Freund's complete adjuvant, induced suppression of murine primary response to SRBC. The suppression could be adoptively transferred with spleen cells depleted of T or B lymphocytes and was best expressed when viable cells were transferred. Some normal rabbit serum has the ability to nullify immunosuppressive effects of the transferred cells.


Anaphylactoid responses of two types of genetically different rats to horse serum and yeast. Rats selectively bred for non-reactivity to clinical dextran (NR rats) fail to exhibit the anaphylactoid reaction to systemic baker's yeast or horse serum. Rats which respond with an anaphylactoid reaction to clinical dextran (R rats) react to baker's yeast but do not react to horse serum. The widespread oedema produced in R rats by systemic baker's yeast provides yet another clear method of differentiating them from NR rats.


Dynamic decision models for clinical diagnosis. A unified approach to clinical decision-making is presented. This combines partially observable Markovian decision processes (Markov or semi-Markov) with cause-effect models as a probabilistic representation of the diagnostic process. Pattern recognition techniques are used in a first stage of system state identification. This new class of dynamic models has a direct application to medical diagnosis and treatment and specific physiological examples are emphasised. The methodology is given for combining the patient state of health, the clinician's state of knowledge of the cause-effect representation from the observation space (measurements), feature selection using pattern recognition techniques and, finally, the treatment decisions with which to restore the patient to a more desirable state of health. A cost functional for the decision process has  then to be optimised according to some pre-assigned objective function (social return from the patient state of health or treatment cost for the patient), when the process has an infinite time horizon.


Epidemiological studies in psychosomatic medicine. The epidemiological triad of host, agent and environment used conceptually in infectious disease may serve as a model for psychosomatic disorders, despite the involvement of many more variables. There are major problems with diagnosis and measurement, however, and the term "psychosomatic" has several meanings. The two main senses are "specific" psychosomatic disorders and an ecological view of illness. The association between psychiatric and physical disorder has been examined in a variety of settings and the findings have suggested that there is a positive relationship. Despite considerable methodological and sampling difficulties in epidemiological research into psychosomatic illness, recent efforts have been made to overcome these. The results of ecological studies appear to be more consistent that those dealing with "specific" psychosomatic disorders and suggest that man has a general psychophysical propensity to disease. Although physical and mental illness do seem to be intimately linked, the reasons for "vulnerability" to illness and "clustering" of illness are obscure. The clarification of these areas appears to be the main task ahead for epidemiology in the field of psychosomatic medicine.


Epidemiological studies of life change and illness. Currently popular methodologies in life changes and illness research are presented in some detail. Selected examples of epidemiological studies utilizing these methodololgies are briefly reviewed. Examples of both retrospective and prospective investigations of life change and illness are given. Discussed in the chapter are the author's views as to limitations of the epidemiologic method and probable directions of future research in efforts to clarify etiologic mechanisms along the pathway between subjects' recent life changes and their subsequent susceptibility to illness.


Psychophysiological responses to the urban environment. The relationship of urbanization to bodily changes and chronic disease is examined. Urban stress, as defined principally by one of its components, crowding, is examined in its role in the development of high blood pressure as well as in the modification of behavior and bodily function. Several sources of data are presented including those from animal studies, observations in man and some new data collected by the authors. The concept of information overload, the effects of noise on performance and on chronic disease, the psychophysiological effects of driving in traffic and the behavioral and bodily effects of crowding in man and animals are all presented. This paper intends to give the reader an overview of the complexity of the relationship of urban stress to behavioral changes and chronic disease. This relationship is described, and its multiple components, both in the environment and the nervous system, are enumerated.


Psychosomatic research today: a clinician's overview. Emphasis is placed on the opportunities and importance, at this time, of continuing with clinically oriented psychosomatic research. For instance, the multidimensional studies of sleep and of depression are beginning to throw new light on psychosomatic processes, and so too are the studies of life events and illness. Short-term prospective clinical investigations provide an attractive framework for such work and they can be complementary to long-term survey studies investigating relationships between constitutional characteristics including personality, and social conditions and disease. There are some useful new tools for measureing psychological characteristics.


Psychiatry and medical progress: therapeutic considerations. Advances in medical technology have spurred interest in psychosocial adaptation to newer procedures, such as cardiac operations, chronic hemodialysis and renal renal transplantation. In spite of this interest, investigators have paid little attention to psychotherapeutic measures involved in the care of patients undergoing these procedures. This presentation reviews the available work in this area and brings together common themes related to the preparation and psychological treatment of these patients. These themes center around the importance of a continuous relationship with the patient which begins in the preoperative and pretreatment phase, the value of exploring in a nonthreatening fashion conflicts and fears aroused by the procedure, the need to counteract the non-human and impersonal aspects associated with medical progress, and the role of psychiatrist as the mediator between the patient and those involved in his care.


Operant conditioning-based behavior modification: one approach to treating somatic disorders. The applicability of behavior modification as a process for treating somatic difficulties is examined within the framework of measurement, modification, and evaluation. Illustrations of this process are presented throughout. In particular, the last section of the paper gives an example of a currently operative and multifaceted approach to the problem of obesity. Among the topics considered herein are the use of behavioral contracts that specify ameliorative weight reduction practices and the assessment as well as manipulation of eating speed.


Psychotropic drugs in somatic disorders. Current trends in the employment of psychotropic medication in somatic illness are considered, and some of the theoretical issues raised by this practice are explored. While much further clinical investigation is needed, a review is made of currently available knowledge regarding psychopharmacotherapy in specific clinical settings: pain, delirium, ischemic heart disease, gastrointestinal disorders, dermatological illness, and in renal, hepatic and pulmonary insufficiency.


Consultation-liaison psychiatry: a psychosomatic service in the general hospital. Consultation psychiatry, an outgrowth of the tenets of psychosomatic medicine that followed the departure from mental-hospital psychiatry to the general-hospital-based psychiatric units, is similar to those consultation services in other specialties available on medical-surgical wards. Its primary aim is to interpret the psychosocial phenomena of health and illness and to redirect the physician from disease- to patient-orientation and to a comprehensive approach. Its functions are diagnostic-therapeutic, instructional and research directed. The psychosomatic orientation is a sine qua non in the understanding of the psychosocial meaning of physical illness, its effect on psychic disability, the psychosocial adjustment of the patient and his family and his ability to adapt to illness and the hospital. An essential feature is the evaluation of the doctor-patient relationship, especially when severe illness requires serious interventions, including the consultation and its reciprocal effect upon the doctor-patient relationship. Liaison activity consists of interpretive-therapeutic mediation between the patient and the therapeutic team when strain endangers their satisfactory cooperation. The therapeutic activity must be short-term and so construed as not to interfere with the primary treatment. The instructional aspect is for the sake of the usual recipients of the service, as well as residents and students. Teaching the interview techniques, especially to students, should demonstrate the holistic approach, comprehensive diagnosis and the need of avoiding orthodox one-sidedness. This paper reviews briefly the scope and role of consultation psychiatry, compares its North American model with its status elsewhere and explores the resistances to it.


Clinical psychosomatic research.

Clinical psychosomatic research. Psychosomatic medicine embrances the influence of psychological factors on the development and course of diverse physical symptoms. Reasons are given why it is recognized that physical morbidity is influenced by such factors, with illustrations from the authors' research on acute ischemic heart disease, peptic ulcer and bronchial asthma. The nature of the psychological factors can be elucidated with the use of recently developed psychometric tests; these relate particularly to mood disturbance, either attributable to affective disorder or prominent personality traits. To insure that samples studied are representative of the disease or symptom being considered, and that all relevant variables are taken into account, the research approach usually necessitates a multidisciplinary team.


Management and care of the dying patient. The recent widespread interest in the problems of the dying patient is reviewed, and the dynamics of the psychological process involved in dying are discussed. Recommendations for the management of the dying patient are presented, with special attention to the issues of 1) truth telling, 2) the essential features patients request themselves in their own care and management, and 3) the unique and individualized goals of treatment.


Electroencephalographic correlates of verbally induced stress in man. This article is a review of some of the sparse investigation of EEG concomitants of verbally induced stress. The difficulties encountered in good experimental design in this formidable area, which may account for the paucity of work, are discussed. A recent experiment in which spectral analysis of EEG played a prominent role, with spectral estimators selected as correlated with stressful questions, is presented in some detail. Autonomic measurements provided the stress indicators. The value of animal experiments for psychiatry in the investigation of stress is briefly examined.


Some problems in the teaching of psychosomatic medicine. The part that social and emotional life plays in the response to stress and the course of illness has been recognized for ages. But the obstacles to the teaching of a "psychosomatic approach" to health and disease persist. This paper explores briefly the attitudinal, definitional, historical and structural factors which present as problems in attempts to teach a psychosomatic approach to medical diagnosis and practice. If there is a body of knowledge rightfully called psychosomatic medicine, and if it is important to appreciate, understand and utilize the psychosomatic approach in everyday clinical work with the whole patient, then new ways must be explored to accomplish this task. Suggestions are offered towards this objective.


Nocturnal psychophysiological correlates of somatic conditions and sleep disorders. Modern sleep research studies have provided the practicing physician with considerable new information concerning the basic psychophysiology of sleep, the effects of medical conditions on sleep and the role of maturational and emotional factors in producing certain sleep disorders. Medical and psychiatric disorders, sleep disorders and drug-induced sleep stage alterations are studied in the sleep laboratory using the same techniques developed to analyze sleep patterns in normal subjects. After initial sleep laboratory adaptation, a profile of the sleep characteristics of various clinical conditions is obtained. This profile can be compared to sleep profiles of normal subjects as well as to the effects on sleep of subsequent experimental or therapeutic procedures. Various studies have shown that coronary artery, duodenal ulcer and nocturnal headache patients experience angina, increased gastric acid secretion and migraine or cluster headaches, respectively during REM sleep. Adult nocturnal asthamtic episodes occur out of all sleep stages while attacks of dyspnea in asthmatic children occur in all stages except stage 4 sleep. Hypothyroid patients show decreases in stages 3 and 4 sleep, while in hyperthyroid patients the percentage of time spent in stages 3 and 4 sleep is markedly increased. Enuretic episodes occur predominantly in non-rapid eye movement (NREM) sleep. Sleepwalking and night terror episodes occur exclusively out of NREM sleep, particularly from stages 3 and 4 sleep. Most child somnambulists and children with night terrors "outgrow" this disorder, suggesting a delayed maturation of the central nervous system. Stimulant drugs are effective in the treatment of the sleep attacks of narcolepsy and in treating certain cases of hypersomnia, while imipramine is an effective treatment for the auxillary symptoms of narcolepsy. Psychological disturbances are frequent in adult somnambulism and night terrors as well as in hypersomnia and insomnia. Proper pharmacologic treatment to provide symptomatic relief for insomnia is recommended to enhance the psychotherapeutic process.


Hypnosis as a treatment method in psychosomatic medicine. Recent research findings are used to illustrate the areas of uncertainty and controversy in the understanding of hypnosis. Despite similarities, hypnosis is presented as more than and different from relaxation, suggestibility, and the placebo response. An overview of the clinical use of hypnosis includes the three main methods of application, namely: relaxation or mild hypnosis, symptom removal, and hypnotherapy. A few brief case reports are included. As a means of narrowing the gap between laboratory research and clinical experience, the author recommends the assessment of hypnotizability in all patients treated with hypnosis.


The relaxation response: psychophysiologic aspects and clinical applications. It is hypothesized that situations requiring continous behavioral adjustment activate an integrated, hypothalamic response, the emergency reaction. The frequent elicitation of the physiologic changes associated with the emergency reaction has been implicated in the development of diseases such as hypertension. Prevention and treatment of these diseases may be through the use of the relaxation response, an integrated hypothalamic response whose physiologic changes appear to be the counterpart of the emergency reaction. This article describes the basic elements of techniques which elicit the relaxation response and discusses the results of clinical investigations which employ the relaxation response as a therapeutic intervention.


Therapy-resistant depressions: biochemical and pharmacological considerations. Two statements are presented here: 1) antidepressants increase the amount of MA at the central receptors, and 2) antidepressants are effective in some, but quite ineffective in other patients, even if they belong to the same diagnostic category. Three questions result from these statements: 1) Does a central MA deficiency occur in depressive patients? 2) If so, is this disorder present in only a proportion of the patients? 3) If so, can this explain the apparent selectivity of antidepressants in the sense that particularly MA-deficient patients benefit from this type of therapy? A tentative answer is given to these three questions. In depressive patients the cerebral MA turnover can be diminished. These disorders do not occur in all patients but seem to be confined to certain categories of depression. Diminution or non-diminution of the central MA turnover is a (not the) factor which determines whether antidepressant medication will succeed or fail. The theoretical and practical implications of these answers are discussed.


DNA breakage, repair and lethality after 125I decay in rec+ and recA strains of Escherichia coli. Iodine-125 decays by electron capture and is known to cause extensive molecular fragmentation via the Augur effect. 125I was incorporated into the DNA of exponentially-growing E. coli K12 AB2487, a recA mutant, and E. coli K12 AB2497, the corresponding rec+ strain, as 5-iododeoxyuridine (IUdR), an analogue of thymidine. Radioactive bacteria were stored at - 196 degrees C, and samples were periodically assayed for loss of viability and for the induction of double-strand breaks (DSBs) in DNA. Each 125I decay in the DNA of either strain induces one DSB, i.e. alpha(DSB) = 1.0. For the recA strain, alpha(lethal) = 0.9 and for the rec+ strain, 0.4. Assays for biological repair of DSBs, involving incubation of thawed samples in growth-medium at 37 degrees C before the extraction of DNA, demonstrate significant repair of 125I-induced DSBs by rec+ cells but none by recA cells. For small numbers of decays, there is approximately a 1:1 correlation, for either strain, between lethal decays and post-incubation residual DSBs. Comparison with data for larger numbers of decays indicates that a typical rec+ cell can repair no more than three to four DSBs per completed genome (2.5 x 10(9) daltons).


Production and rejoining of single-strand breaks in DNA of Escherichia coli cells after exposure to gamma-rays in the presence of iodoacetic acid under oxic and anoxic conditions. The presence of iodoacetic acid (IAA, 10(4) M) during exposure of Escherichia coli cells to 25 krad gamma-radiation under anoxia resulted in a greater number of radiation-induced single-strand scissions in DNA. The single-strand breaks produced under oxic conditions of irradiation, however, were only marginally increased when IAA was present during irradiation. Although cells irradiated under anoxic or oxic condition and those irradiated under anoxic condition in the presence of IAA were capable of rejoining single-strand breaks produced in DNA, those irradiated under oxic condition in the presence of IAA had lost this capacity.


Lethality and double-strand scissions from 14C decay in the DNA of micro-organisms. 14C-2-thymidine was incorporated into the DNA of E. coli B/r and of coliphage T4. The labelled organisms were stored for several years at -196 degrees C. Both were periodically assayed for loss of viability, and the coliphage also for the appearance of double-strand breaks (DSBs) in DNA. E. coli B/r exhibited a survival curve with a substantial initial shoulder, extrapolation number 5-2 +/- 2-3, and a final exponential portion corresponding to a lethal efficiency per 14C decay per 2-5 X 10(9) daltons of DNA, of 0-009 +/- 0-002. For coliphage T4, our best estimate for the lethal efficiency per 14C decay is 0-03 +/- 0-04, and that for the DNA breakage efficiency is -0-002 +/- 0-004. The large standard errors result from the very small number of 14C decays occurring in each phage. These results suggest that 14C decay in the DNA of micro-organisms does not cause DSBs but does cause potentially lethal damage to the thymine bases in which decay occurs, and that wild-type E. coli can repair a large number of such DNA lesions.


Medical and surgical management of corneal thinnings and perforations. The primary objective of treatment of a thin cornea is to maintain the integrity of the anterior chamber and to control the ocular inflammation. Available methods of therapy range from simple medical means, through soft contact lenses and tarsorrhaphy, to conjunctival flaps and keratoplasty. Once perforation has occurred, if the anterior chamber has not reformed in two to three days despite patching, soft contact lenses or adhesives should be tried; if not successful, the more complicated procedures such as glued-on hard contact lens, a blow-out patch graft, or penetrating keratoplasty must be undertaken to restore the integrity of the eye.


Penetrating keratoplasty for herpetic keratitis: decision-making and management. The outlook for keratoplasty in herpetic keratitis has improved significantly over the past decade. While only two out of three grafts in vascularized and regrafted eyes and in eyes with active disease are clear after two years, in quiet eyes with no significant vascularization, the success rate exceeds 80 percent. The significant factor in the improved results, in both favorable and unfavorable cases, is the improvement in surgical technique, involving the operating microscope, the 10-0 nylon suture, and the increased understanding of the pathophysiology of corneal grafting, which has resulted in more sophisticated postoperative care. In our experience, the routine use of high levels of topical steroids in the immediate postoperative period, followed by a rapid tapering of these medications, has been particularly successful.


Rubella.

Rubella. Experience with the vaccine can be summarized as follows: There were minimal problems with neuralgia and neuropathy as a result of the vaccine; vaccinated children were not a significant source of infection to the pregnant woman; the duration of immunity is similar to that obtained with natural rubella virus, but with lower antibody levels; and reinfection is more common with vaccination, but apparently no viremia develops. Several studies were reported regarding inadvertent vaccination in women who were pregnant at the time of vaccination. In one series [7], 19 pregnant women who were vaccinated were studied. Ten went to term and delivered normal babies and 9 aborted, 8 induced and 1 spontaneously. In one induced abortion, virus was demonstrable in fetal tissue including, notably, in the eye. Another case of CRS was diagnosed in utero amniocentesis [12], and the fetus was aborted at 12 weeks' gestation. Another evaluation of reinfection during pregnancy [1] confirmed that this was of little risk to the fetus, as there was no apparent viremia. In summary, then, in one ophthalmological generation, from Gregg until the present time, we have experienced the recognition, the observation, and, essentially, the conquest of the congenital rubella syndrome. However, if vaccination programs are not assiduously continued, we can expect a renewed upsurge of this difficult problem.


Surgical management of vertebral-basilar insufficiency. The authors report treatment of 251 lesions of the major branches of the aortic arch and the vertebral artery in 172 patients. A detailed account is given of the lesions observed (34 lesions of the innominate artery, 93 of the subclavian artery and 119 of the vertebral arteries), the technics used and the indications for surgery. Use of the cervical approach seems to be justified in order to adapt the extent of surgery to the patient's condition. This accounts for the low mortality rate of 1.5%.


[Findings and significance of spermatozoa agglomeration in the semen]. Agglomeration of spermatozoa (pseudo-autagglutination, clumping of spermatozoa) is distinguished from agglutination in immunological processes by the coarse meshwork of the spermatozoa clumps and the central inclusion of epithelia, leukocytes, residual bodies, spermatozoa with persisting cytoplasmatic droplets, as well as parts of viscous seminal plasma. In 658 semen samples of out-patients with barried marriage, spermatozoa agglomerations were seen in 13.4% of the samples immediately after liquefaction while in additional 6.7%, they were found up to three hours later. The agglomeration phenomenon is typical but not specific of the so-called vegetative congestion syndrome, appearing concomitant with high spermatozoa counts, normal rate of spermatozoa motility and morphology, as well as an increase in residual bodies and spermatozoa with persisting cytoplasmatic droplets. Next to congestion syndrome, spermatozoa agglomerations may also result from post-inflammatory disturbances of epididymal or accessory gland function whereas in florid stages of inflammation, they are rarely observed. As the congestion syndrome may give rise to male infertility in some cases, andrologists should be acquainted with its clinical and spermatological symptoms with special regard to therapy.


Mycoplasma and L-forms: occurrence in bacterial cultures. The mycoplasma and the L-forms of bacteria have been associated since the time of their discovery. Their similar biological properties have raised questions concerning their common origin. Observations made over a number of years have shown the appearance of mycoplasma in L-form cultures of several bacterial species. The origin of these mycoplasma cultures remains unknown. Recent research suggests the possibility of phylogenetic relationships between mycoplasma and bacteria or their derived L-forms.


[Therapy of facial hemangioma].

[Therapy of facial hemangioma]. Indication, advantages and disadvantages of 5 methods for the treatment of hemangiomas of the face are discussed.


[100 consecutive colonic resections with single layer sutures and early radiologic postoperative controls]. A prospective study concerning 100 consecutive cases of colonic resections and anastomosis with single layer sutures is reported where radiological controls 10-12 days postoperatively had been carried out. An anastomotic insufficency was recognized clinically in 15%; in 6% a small leakage was only detectable radiologically. 50% of all anastomotic deficencies were seen in cases with preexisting perforation peritonitis or ileus, especially if an emergency resection had been necessary due to septic complications. 3 out of the 7 deaths were connected with dehiscence in the suture line.


Inhibition of plaque growth with taurolin, vantocil and amine fluoride. 16 dental students rinsed daily with a placebo solution, Taurolin 2% (formaldehyde releasing antimicrobial), Vantocil 0.1% (polybiguanide), and with amine fluoride 0.025%. Depression of plaque growth assess with the Sulcular Plaque Index was 50.6, 40.2 and 21.5%, respectively.


Cholorate toxicity in Aspergillus nidulans: the selection and characterisation of chlorate resistant mutants. Mutation in at least ten genes can result in chlorate reistance in Aspergillus nidulans. Mutation in seven of these genes also results in the inability to use nitrate as nitrogen source. The various classes of resistant mutant obtained occur in different proportions, depending on whether or not a mutagenic treatment is employed, and also on which nitrogen source is used for selection. The prinicipal effect of mutagen arises because mutations in the niaD gene, the nitrate reductase structural gene, are relatively much commoner when no mutagen is used than after treatment with N-methyl-N'-nitro-N-nitrosoguanidine. This may be connected with the finding that deletions involving the niaD gene  are relatively more common among samples of spontaneous niaD mutants. Some of these deletions extend to the neighbouring niiA gene, the structural gene for nitrite reductase. The selection procedures used were designed to avoid bias in favour of any particular chlorate resistant phenotype. Even if biases existed however, these could not account for the variation found from nitrogren source to nitrogen source in the proportions of certain resistant classes having apparently identical chlorate resistance phenotypes.


The variable refractivity of the protein or polypeptide hormone-producing cells showing a unique luminescence in the dark-field microscope. When cryostat sections of endocrine tissue were examined in a dark-field microscope, a brillant granular luminescence was revealed in the endocrine cells thought to be concerned with protein or polypeptide horome production. The sections were prepared from fresh materials either frozen in a cryostat chamber at --25 degrees C, in dry ice-acetone, or fixed in formalin-calcium for 24 hr. The neurosecretory substance in the hypothalamus and the posterior lobe of the pituitary showed a blue luminescence; the acidophil cells of the anterior lobe of the pituitary, orange; basophile cell, green or blue; intermediate lobe cells, no luminescence; thyroid C cells, white-blue; pancreatic A cells, blue; B cells, orange; adrenomedullary cells, greenish blue; enterochromatin cells, green; and other endocrine cells in the gastrointestinal tract, blue or orange. After tearing and spreading the pituitary and hypothalamus with a pair of needles on a glass slide, and examining the teased specimen by dark-field microscopy, various cells of different lumunescent colours became apparent in the anterior lobe of the pituitary, a blue fluorescent substance in the posterior lobe, and neurosecretory cells bodies in the hypothalamus. The different colours appear to be inherent in the granules of living tissues.


Origin and early evolution of transition element enzymes. In this paper we speculate on the origin and early evolution of transition element enzymes. Iron, molybdenum, and zinc, the most abundant transition elements in seawater, presumably complexed with compounds accumulated in the primeval sea in the course of chemical evolution forming compounds with subsequently evolved to form proenzymes or early enzymes with low activity and broad specificity. Iron complexes may be regarded as precursors of electron transfer enzymes, molybdenum complexes as precursors of enzymes involved in the metabolism of small molecules, and zinc complexes as precursors of hydrolytic and transferring enzymes, including enzymes participating in the metabolism of macromolecules and information transfer. The different iron, molybdenum, and zinc enzymes found in bacteria including Clostridium may then have arisen through specialization by increases in the enzyme specificity of these proenzymes. Copper would have been incorporated as an enzyme constituent after the elevation of environmental redox potential, probably due to the accumulation of atmospheric oxygen.


R17 RNA replicase. V. Rifamycin sensitivity of R17 RNA synthesis in vivo. The effect of rifamycin on R14 phage growth in vivo was examined using rifamycin sensitive and resistant strains of E. coli as host cells. The following conclusions were obtained: (1) There is a time lag between the addition of rifamycin to the culture and the initiation of inhibitory action by the drug. At 25 mug rifamycin per ml, this time lag is approximately 30 min. At 50 mug per ml, it takes 10 min to exert 90% inhibition of RNA synthesis in both infected and non-infected cultures. (2) The rifamycin sensitive stage for R17 growth is the first 20 min of its infectious cycle during which time the synthesis of phage components, but not the assembly, takes place. (3) Of the phage component synthesis, RNA synthesis is definitely sensitive to rifamycin in contrast to the other RNA phage systems. Of the four phage specific RNA's, progeny plus strand synthesis is trongly inhibited. The synthesis of replicating forms is also sensitive to rifamycin, but their suppression appears to be incomplete.


R17 RNA replicase. VI. A large scale preparation of R17 template RNA. A new procedure for R17 RNA preparation was devised using a 300 liters fermenter. The key factor in processing such a large quantity is the purification of phage particles prior to RNA extraction. The method involves the preparation of 250 liters of crude lysate, condensation of phage particles by the partition method, purification by DEAE-cellulose column, and removal of adherent proteins by a series of high-salt washes. The method permits preparation of approximately 3 g of phage particles free of ribosomal fragments and RNase. Phage RNA extracted in gram quantities using conventional methods often contain phenol. Thus repeated extraction of R17 RNA with salt-alcohol mixture is required.


Fluorescence polarization studies on the local conformation of yeast tRNA. The effects of temperature and guanidine hydrochloride (GuHC1) on the fluorescence polarization of Y base in partially purified yeast phenylalanine tRNA (p.p.tRNAPhe) and in acriflavine conjugates of crude yeast tRNA were investigated to elucidate the stability of the conformation at the 3'-CpCpA terminus and the anticodon loop of tRNA. The results can be summarized as follows: (1) The kinetic unit of the internal rotational motion near Y base in the anticodon loop of p.p.tRNAPhe seems to be independent of the temperature over the range 5--60degrees. Accelerated depolarization is observed in the 3'-CpCpA terminus at temperatures over 30degrees. This suggests that a change occurs in the native conformation of the 3'-CpCpA-acriflavine terminus at temperatures over 30degrees. (2) The fluorescence polarization of Y base in p.p.t-RNAPhe is not affected by the addition of GuHC1 up to 6 M, while that of acriflavine conjugated to the 3'-CpCpA terminus decreases markedly in the presence of 0.8 M GuHC1 reflecting the disruption of the native conformation. These results indicate that the local conformation near Y base is stable, but that of the amino acid acceptor terminus is not.


Studies on ribosomal ribonucleic acid from yeast. III. Secondary structure of 18 and 26S yeast ribosomal RNA's and their complex: circular dichroism and infrared analyses. Circular dichroism (CD) and infrared (IR) spectroscopic analyses were performed of 26 and 18S yeast ribosomal RNA's (rRNA) and their specific complex, 30S RNA. The molecular ellipticity coefficients [thota] of 18, 26, and 30S rRNA's were 2.72, 2.63, and 2.65X10(4) degree-cm2/decimole at 264 nm, respectively. The base-pairing contents of 18, 26, and 30S rna's determined by iC), and 70% (32% AU, 38% GC), respectively. These results suggest that 18 and 26S rRNA have very similar secondary structures, and that 30S rna may have a slightly higher base-pairing content than the estimated sum of those of 18 and 26S rRNA's. The biological significance of this phenomenon is discussed in this report.


Effects of asparagusate and lipoate on enzymes of the tricarboxylic acid cycle and related metabolic pathways. 1. The effects of lipoate and asparagusate on animal and plant enzymes of the TCA cycle and related metabolic pathways were studied. 2. Lipoate inhibited bovine liver glutamate dehydrogenase [EC 1.4.1.3]. The inhibition may play a role in metabolic regulation. 3. Asparagusate inhibited lipoyl dehydrogenase [EC 1.6.4.3] from asparagus and lettuce competitively with respect to lipoate. Asparagusate had practically no effects on other asparagus enzymes. 4. Asparagusate strongly inhibited lipoyl dehydrogenase, glutamate dehydrogenase, and isocitrate dehydrogenase [EC 1.1.1.42] from animal sources, in competition with the corresponding substrate. 5. Asparagusate and lipoate also inhibited yeast glutamate dehydrogenase. 6. Based upon kinetic studies, the mode of these inhibitions is discussed.


Peptic peptides from the beta polypeptide chain of AII component of chicken hemoglobin. The aminoethylated beta polypeptide chain of AII component from chicken hemoglobin was digested with pepsin [EC 3.4.23.1] and the resulting peptides were separated and purified by gel filtration, ion exchange chromatography, and paper chromatography. The amino acid composition and partial sequence of the peptic peptides were studied. From the results thus obtained, the primary structure of the beta polypeptide chain was established, taking account of the amino acid sequences of the tryptic peptides previously reported.


Application of the enzymic electric cell method to the activity assay of NAD-linked dehydrogenases. An enzymic electric cell was constructed with a saturated calomel electrode (cathode) and an enzymic electrode (anode) which consisted of a glassy carbon electrode and a mixture containing an NAD-linked dehydrogenase, NAD+, Nomethylphenazonium methosulfate and a substrate, and the short-circuit current of the cell was measured...


Dessociation and reconstitution of colicin E3 and immunity substance complex. It has recently been found that so-called native colicin E3, which has been used for studies of its mode of action, is a complex of two kinds of proteins. The complex could be dissociated into the two components in SDS. These components were isolated by gel filtration of 1% SDS followed by treatment with Sowex-2 to remove bounds SDS. One component, characterized by its low molecular weight, prevented colicin E3-induced inhibition of poly(U)-dependent protein synthesis and was designated as immunity substance. The other component (protein A), which was of high molecular weight, had 100-fold higher in vitro ribosome-inactivating activity than native colicin E3, but had lower bacteriocidal activity. Colicin E3 was reconstituted from the two isolated protein components. The reconstituted colicin E3, when compared with protein A, showed a decrease in in vitro activity (inhibition of poly(U)-dependent protein synthesis), but had higher bacteriocidal activity in vivo. Thus complex formation of protein A with immunity substance should play and important role in the bacteriocidal action, but protein A itself might inactivate ribosomes in the interior of the sensitive cells.


Endonuclease II of Escherichia coli is exonuclease III. Exonuclease III, a phosphatase-exonuclease specific for bihelical DNA, wn the preparation was endonuclease II, an activity specific for DNA that has been partially depurinated by treatment with methyl methanesulfonate. The two activities, which could not be separated by electrophoresis, by sedimentation, or by gel filtration, were associated with a single monomeric protein of 28,000 daltons. To explain how a relatively small protein could have such diverse activities, it is proposed that one site on the enzyme can recognize interstrand spaces created either by depurination or by spontaneous terminal unwinding of a DNA duplex.


Site-specific interaction of Qbeta host factor and ribosomal protein S1 with Qbeta and R17 bacteriophage RNAs. We have studied the interaction of the host factor (HF) required for bacteriophage Qbeta RNA replication and of ribosomal protein S1, a subunit of Qbeta replicase, with Qbeta and R17 RNA. Both proteins bind to both Qbeta and R17 RNA; HF has a higher affinity than S1 for these phages RNAs. HF binds to a single site in R17 RNA located in the replicase cistron, and to two sites of Qbeta RNA, one of which is located approximately 60 nucleotides from the 6' end of Qbeta RNA. The three HF binding sites all have portions rich in adenylate residues; all are bound by HF when contained in oligonucleotides which are predicted to exist only in single-stranded form. S1 selects a single site in Qbeta RNA, also near the 6' end, but binds to a large number of sites in R17 RNA. These results suggest that HF and possibly S1, through their interaction with the 3'-terminal region of Qbeta RNA, are directly involved in the recognition of the 6' end of Qbeta RNA by Qbeta replicase. Under conditions where specific protein-R1M RNA complexes are formed, we have also tested host factor and S1 for cistron-specific interference with ribosome binding to R17 RNA. Although S1 and HF lower the efficiency of initiation complex formation as described previously, we detect no discrimination against any particular cistron. We therefore conclude that translational interference exhibited by the two proteins probably reflects simply their high affinity for RNA and certain defined polynucleotides.


Effect of estrogen on gene expression in the chick oviduct. Hen oviduct RNA polymerase II and Escherichia coli RNA polymerase holoenzyme and core enzyme were used to study the initiation of RNA synthesis on chromatin. In either the presence or absence of estrogenic stimulation, changes in the level of oviduct chromatin initiation sites as measured in the presence of either homologous or heterologous polymerases followed a similar pattern. Comparison of the initiation sited utilized by these enzymes on chick oviduct chromatin indicated that these enzymes compete with each other for the same initiation regions. In contrast to chromatin, however, the majority of the initiation sites on DNA which are utilized by the oviduct RNA polymerase II are different from those utilized by E. coli holoenzyem. These results suggest that chromatin proteins are involved in the selection of initiation sites on chromatin for RNA polymerases. The in vitro transcripts of these RNA polymerases on stimulated chick oviduct chromatin were analyzed by hybridization to a cDNA probe transcribed from ovalbumin mRNA. The relative concentration of ovalbumin sequences transcribed by these three polymerases was 4:1.5:1 for oviduct RNA polymerase II, E. coli core enzyme, and holoenzyme respectively. Therefore, the efficiency of transcribing a specific gene appears to depend on the interaction between RNA polymerase and chromosomal elements in the initiation region.


A reaction mechanism from steady state kinetic studies for O-acetylserine sulfhydrylase from Salmonella typhimurium LT-2. It has been determined from steady state kinetic studies using the sulfide ion selective electrode that O-acetylserine sulfhydrylase catalyzes a Bi Bi Ping Pong reaction between O-acetyl-L-serine and sulfide. Both O-acetyl-L-serine (OAS) and sulfide exhibit strong competitive substrate inhibition. A fit of all the data to the equation for the mechanism yields KOAS = 0.149 +/- 0.059 mM and KIOAS = 46.91 +/- 10.06 mM for O-acetyl-L-serine and KS2- = 0.066 +/- 0.004 mM and KIS2- = 0.013 +/- 0.006 mM for sulfide. Product inhibition studies varying either substrate at changing fixed levels of cysteine demonstrate that cysteine combines with enzyme at two places along the reaction sequence to produce inhibition with KiCys = 1.048 +/- 0.048 mM and KICys = 11.4 +/- 0.5 mM. Relatively high concentrations of acetate are required to produce inhibition and at least part of the acetate inhibition is due to ionic strength. However, the ability of acetate to reverse the spectral shift produced from the binding of O-acetyl-L-serine to enzyme and the isotope exchange between [14C]acetate and O-acetyl-L-serine does demonstrate that the O-acetyl-L-serine to acetate half-reaction is reversible. There is some doubt as to the specificity of acetate as a product inhibitor, since propionate can also be used to reverse the spectral shift. Spectral studies using ths spectral shift produced from binding O-acetyl-L-serine to enzyme confirms the assignment of a ping-pong mechanism since the spectral intermediate produced is alpha-aminoacrylic acid in Schiff base with pyridoxal phosphate and, therefore, the acetyl moiety has been beta eliminated. Isotope exchange has been demonstrated for both the O-acetyl-L-serine to acetate and sulfide to cysteine half-reactions which also confirms a ping-pong mechanism.


Characterization of DNA polymerase induced by bacteriophage T5 with DNA containing single strand breaks. DNA polymerase induced by bacteriophage T5 was purified and characterized using mainly circular duplex DNA of bacteriophage PM2 with single strand breaks formed by DNase I action. A purification procedure is described which has consistently yielded DNA polymerase preparations with only one detectable protein band after polyacrylamide gel electrophoresis of either native protein in Tris-glyase preparations utilized both denatured DNA and nicked DNA as primer-templates, although at 37 degrees the activity with denatured DNA was much greater. Polymerase activities with both kinds of primer-templates were shown to be associated with one phage-induced protein. DNA synthesis with nicked DNA as primer-template increased with increasing numbers of single strand breaks. Essentially all such breaks were repairable by ligase. Alkaline sucrose gradient centrifugation showed that synthesis occurred with the strand which had a single strand break as a primer yielding DNA longer than one phage DNA unit length. Newly synthesized DNA was covalently linked to the primer strand. Thus the synthesis very likely occurred by strand displacement; this is supported by electron micrographs shown in the Appendix.


Outer membrane of Salmonella. Isolation of protein complex that produces transmembrane channels. Salmonella typhimurium outer membrane protein complexes can be reconstituted with lipopolysaccharide and phospholipids into membrane vesicles. These vesicles are permeable to a variety of low molecular weight compounds, but not to oligo- and polysaccharides of molecular weight higher than 700. A protein complex participating in selective membrane permeability can be isolated by gel filtration in the presence of sodium dodecyl sulfate. The active fractions contain three major protein species. The Braun lipoprotein is not associated with this subset of outer membrane proteins.


Methylated, blocked 5' termini of yeast mRNA. mRNAs of the yeast, Saccharomyces cerevisiae, have modified 5' termini similar but not identical with those present in the higher eukaryotes. Poly(A)-containing RNA isolated from spheroplasts or from polysomes of cells labeled with [methyl-3H]methoionine or 32Pi contain methylated, blocked 5'-terminal structures of composition: m7G(5')pppAp and m7G(5')pppGp, occurring in a relative distribution of 75% and 25%, respectively. Furthermore there is no N6-methyladenosine nor any other methylated nucleoside in the entire molecule of poly(A)-containing RNA. Experiments with a mutant, temperature-sensitive for the transport of mRNA to cytoplasm, suggest that the methylation and addition of the blocking group take place in the nucleus and that the genetic lesion in the mutant does not affect these mechanisms.


Subunit and chemical composition of citrate lyase from Klebsiella pneumoniae. Citrate lyase from Klebsiella pneumoniae (previously called Klebsiella aerogenes) is shown by amino acid sequencing techniques to contain equimolar amounts of three nonidentical subunits. Together with the molecular weight of the enzyme, this finding is interpreted to indicate that there are 6 mol of each of these subunits per mol of native enzyme. A partial amino acid sequence of each of the three subunits is presented. The pure enzyme with a specific activity of 73 units/mg of dry protein has an absorbance index (E1% 1 cm at 278 nm) of 6.2. The amino acid composition of the native enzyme is presented. In addition, the amounts of adenine, phosphate, taurine (from cysteamine), and "ribose," present as a bound cofactor of unknown structure, have been estimated. A comparison of amino acid compositions and the partial amino acid sequences of two of the subunits reveals some resemblance between them.


Synthesis of human placental lactogen and human chorionic gonadotropin by polyribosomes and messenger RNA's from early and full term placentas. Synthesis of human placental lactogen (hPL) and of human chorionic gonadotropin (hCG) by membrane-bound and free polyribosomes from early and from full term human placentas was investigated by in vitro release of the nascent hormone peptides, followed by immunoprecipitation and electrophoresis in sodium dodecyl sulfate gels, and by specific binding of 125I-labeled hPL antibody to nascent peptide chains. In addition, messenger RNA'S Were extracted from total, free, and membrane-bound placental polyribosomes and their capacities for hPL and hCG synthesis were measured in a heterologous cell-free system prepared from wheat germ. Membrane-bound polyribosomes from full term placentas were several times more active in the synthesis of both peptide hormones than were free polyribosomes. By binding 125I-labeled hPL antibody to nascent chains on the polyribosomes, it was determined that hPL is made by clusters of seven to nine ribosomes. About 8% of the nascent peptide chains released by incubation of polyribosomes from full term placentas was accounted for by hPL, and 2% by hCG. In contrast, no chains of hPL were released by polyribosomes from 10-week placentas, whereas 11% of the total released chains were accounted for by hCG. When messenger RNAs prepared from the polyribosomes of 20- and 40-week placentas were used to stimulate protein synthesis in a wheat germ system, hPL accounted for 0.4 and 2%, respectively, of total protein synthesis, while hCG was 8 and 2%, respectively. This confirmed the relative proportions observed for nascent chains on the polyribosomes of early and late placentas. Unexpectedly, translation of mRNA from free polyribosomes yielded as much hPL and two-thirds as much hCG as did translation of mRNA from bound polyribosomes. We conclude, that the decreased blood levels of hCG and increased blood levels of hPL with advancing gestational age reflect the relative in vitro rates of synthesis of these hormones by placental polyribosomes, the abundance of which is determined by availability of their respective messenger RNAs at different times in gestation.


Roles of individual mgl gene products in the beta-methylgalactoside transport system of Escherichia coli K12. Previous findings showed that galactose-binding protein defective mutants (mgl B-,A+,C+) of Escherichia coli K12 are still capable of growth on methyl-beta-D-galactopyranoside, while mgl A- and mgl C- mutants are not. When assayed by previous methods, none of these mutants exhibited methylgalactoside transport system activity. In this study, we present a modified assay developed for measuring low levels of transport. Using this assay, we found that mgl B-,A+,C+ mutants defective in galactose-binding protein accumulate methyl-beta-D-galactopyranoside up to six times the concentration gradient while mgl A- and mgl C- mutants failed to accumulate this substrate. Similar results were obtained using D-glyceryl-beta-D-galactopyranoside, another substrate of the methylgalactoside transport system. In contrast, all sugars tested which are not substrates of this system were transported equally by all mgl- mutants. The kinetic parameters of transport in mgl B- mutants were compared to those of the isogenic mgl+ strain which accumulates methyl-beta-D-galactopyranoside against a 10,000-fold concentration gradient. The apparent Km of methyl-beta-D-galactopyranoside influx was 1,000 times greater in mgl B- than in mgl+ strains. In contrast, there was no significant difference between these strains in either the Vmax of substrate influx or the rate of substrate exit. D-Galactose competitively inhibited methyl-beta-D-galactopyranoside influx into both mgl B- and mgl+ strains; the Ki of inhibition in mgl B- cells was 2,000-fold greater than that in mgl+ cells.


Dead or alive: the problem of in vitro tissue mechanics. Extensive studies have been made of the biomechanical properties of tissues in vitro. The majority of experiments demonstrate a "preconditioning" effect. This effect is associated with the differences between in vivo and in vitro conditions. Failure to utilize initial or isochronal data may lead to erroneous conclusions. Objective criteria would permit comparison between experiments.


Maintenance of aseptic barriers in the conventional operating room: general principles. Contamination of operative wounds during surgery occurs primarily through contact and secondarily through the airborne route. Inspection and maintenance of air-handling equipment is essential, with special reference to air intakes, filters, humidifiers, and exhaust systems. Doors should remain closed to maintain positive pressure. Surgical attire for all personnel must provide for complete coverage of hair and arms. Intensive interim cleaning of all horizontal sufaces between operations is essential. The infection rate is proportional to the duration of the operation and the number of personnel in the room, and inversely proportional to the air changes per hour.


Prevention of infection in the treatment of one thousand and twenty-five open fractures of long bones: retrospective and prospective analyses. In 673 open fractures of long bones (tibia and fibula, femur, radius and ulna, and humerus) treated from 1955 to 1968 at Hennepin County Medical Center, Minneapolis, Minnesota, and analyzed retrospectively, the infection rate was 12 per cent from 1955 to 1960 and 5 per cent from 1961 to 1968. In a prospective study from 1969 to 1973, 352 patients were managed as follows: débridement and copious irrigation, primary closure for Type I and II fractures and secondary closure for Type III fractures, no primary internal fixation except in the presence of associated vascular injuries, cultures of all wounds, and oxacillin-ampicillin before surgery and for three days postoperatively. In 158 of the patients in the prospective study the initial wound cultures revealed bacterial growth in 70.3 per cent and the infection rate was 2.5 per cent. Sensitivity studies suggested that cephalosporin is currently the prophylactic antibiotic of choice. For the Type III open fractures (severe soft-tissue injury, segmental fracture, or traumatic amputation), the infection rates were 44 per cent in the retrospective study and 9 per cent in the prospective study.


A highly ordered ring of membrane-associated filaments in budding yeast. In Saccharomyces cerevisiae, a highly ordered ring of 10-nm filaments is intimately associated with the plasma membrane within the neck of the bud. The ring is formed during early bud emergence and disappears when cytokinesis begins.


Glucagon metabolism in man, studies on the metabolic clearance rate and the plasma acute disappearance time of glucagon in normal and diabetic subjects. To investigate further the "hyperglucagonaemia" of diabetes mellitus, the metabolic clearance rate (MCR) and acute disappearance time (t1/2) of unlabelled pancreatic glucagon were estimated in 9 normal subjects and 7 insulin-dependent diabetics, using a constant infusion technique. The mean MCR (+/-SE) was similar for both groups (control: 9.0 +/- 0.6; diabetics: 11.4 +/- 1.0 ml/kg/min). The MCR was not influenced by the concentration of glucagon at the time of plateau, and the exogenous hormone appeared to be handled similarly to endogenous glucagon. On the other hand, t1/2 calculated from the fractional decay rate of glucagon from plateau was significantly prolonged in the diabetics (t1/2 6.6 +/- 0.5 min) compared with the control group (t1/2 4.8 +/- 0.2 min, P less than 0.01). Furthermore, there was no correlation between MCR and t1/2, for the control, diabetic, or combined group. It therefore appears that the MCR of glucagon is similar in normal and diabetic subjects. However, since the acute disappearance time (t1/2) of glucagon is not identical in these two groups, it appears that the kinetics of the overall in vivo metabolism of pancreatic glucagon are not similar in diabetic and control subjects.


Surgical management of vertebral-basilar insufficiency. The authors report 251 cases of lesions of the major branches of the aortic arch and the vertebral artery in 172 patients. A detailed account is given of the observed lesions (34 lesions of the innominate artery, 93 of the subclavian artery and 119 of the vertebral arteries), the techniques used and the indications for surgery. The use of the cervical approach seems to be justified in order to adapt the extent of cure to the patient's condition and accounts for the low mortality rate of 1.5%.


Effects of somatostatin on basal levels of plasma growth hormone and insulin in acromegalics: dose-response studies and attempted total growth hormone suppression. Somatostatin 5, 10, and 25 mug and saline were given as a 2 min bolus injection to 6 acromegalic patients in basal conditions. A significant dose-response relationship could be demonstrated between somatostatin and the suppressions of plasma growth hormone and insulin. The lowest somatostatin dose tested exerted a significant suppression of both hormones. Insulin suppression after the bolus injection lasted for 15 minutes, while the suppression of growth hormone was maintained for 30-50 minutes. In order to obtain total suppression of the elevated plasma growth hormone levels in acromegalics, 3 patients received 50, 250, and 500 mug of somatostatin as bolus injections at time 0, 90, and 180 minutes, 2 patients received 50 mug of somatostatin as bolus injections 4 times with an interval of 20 minutes between each injection, and finally 3 patients received a large dose of somatostatin, 3000 mug given as an infusion over 2 hours. The single injections of somatostatin were not followed by a satisfactory growth hormone suppression. In the infusion experiments, the average plasma growth hormone level was suppressed only 65%, resulting in individual plasma growth hormone plateaus of 20, 14, and 3.6 ng/ml. Only the lowest of these plateaus would be acceptable from a clinical point of view.


Human mixed somatotrophic and lactotrophic pituitary adenomas. Six patients with acromegaly at examination were found to have pituitary adenomas composed of cells that secreted GH and PRL. This was demonstrated by the elevated serum hormone concentrations, by immunoperoxidase staining of 5 specimens, and by electron microscopic examination of 4. Ultrastructural characteristics, described in detail, suggest that these adenomas were mixed adenomas consisting of 2 well-defined, distinct cell types, each secreting one hormone. By immunoperoxidase staining some cells were found to contain immunoreactive growth hormone, other cells immunoreactive prolactin. No cells were detected exhibiting immunostaining for both growth hormone and prolactin. Eelctron microscopy, consistent with the results of immunostaining, revealed the presence of two distinct cell types, distinguishable from each other by their characteristic fine structural features. No intermediate forms were noted. Thus there was no evidence to suggest that one cell type might transform  to the other. Present findings seem to indicate that mixed adenomas secreting growth hormone as well as prolactin and consisting of somatotrophs as well as lactotrophs do occur in the human pituitary gland. Although all the results obtained so far suggest that these tumors are composed of two distinct cell types and thus can be interpreted as representing real mixed adenomas, further work is required to establish whether or not they derive from one common progenitor.


Dexamethasone suppression of serum T3 and T4. Dexamethasone (2 mg q6h for 48 h) decreased serum T3 in normal and athyreotic subjects, and decreased T4 in normal subjects. Dexamethasone probably alters secretion and peripheral metabolism of thyroid hormones.


Diet-induced alterations of hGH secretion in man. Studies were designed to determine whether variations in diet composition could modify the secretion of human growth hormone. Eight men and seven women ingested experimental diets for 10-12 days. Each experimental diet was preceded by a control diet for five days. Experimental diets studied in men were a) 2300 calorie, 80% carbohydrate (8 men); b) 2300 calorie, 75% high-fat (7 men); c) 2300 calorie, 70% high-protein (5 men); d) 3600 calorie, "control" (40% carbohydrate, 40% fat, 20% protein) (5 men); and e) 3600 calorie, 80% high-carbohydrate (5 men). A control diet and a high-carbohydrate (5 men). A control diet and a high-carbohydrate diet at the 2300 calorie level were studied in women. Each diet study was terminated by a 72 hour fast. Serum samples were collected hourly for 24 hours after each control period, on the eigth, ninth, or tenth day of each study, and during the final day of each fast. High-carbohydrate diets at the 2300 calorie level caused a significant decrease of growth hormone values in serum in each of eight men (sign test of significance, P less than .01). The mean figures were likewise significantly decreased. Isocaloric diets of high fat and high protein did not alter growth hormone concentrations in serum. A high-caloric diet similar to the control diet in composition was without effect on growth hormone secretion in men; however, a high-carbohydrate diet at the higher caloric level again depressed growth hormone values in plasma. On the third day of a 72 hour fast, growth hormone values in serum increased 287% in men, from a mean control serum concentration of 4.4 +/- 0.8 ng/ml to 11.9 +/- 5.0 ng/ml (P less than .01). Women, unlike men, had no significant decrease in growth hormone concentrations in serum over a 24 hour period after the high-carbohydrate diet, and the increase after starvation was significantly less than that in men, achieving significance only when evaluated by paired analysis. Growth hormone values in serum after the infusion of arginine followed a similar pattern, i.e., decreased after high carbohydrate but unaffected by other diets in men; high carbohydrate diets did not alter the growth hormone response of women to arginine.


Recovery of yeast from vented blood culture bottles. Rates of isolation of yeasts from blood cultures were significantly enhanced by venting vacuum blood culture bottles in studies of both stimulated and patients' blood cultures; however, the time interval to detection of positivity of yeasts in the clinical studies was significantly (P less than 0.01) shorter in a vented bottle with biphasic brain heart infusion medium than in a vented bottle with soybean-casein digest broth. The mean time intervals to detection of positivity were 2.6 days in the former and 5.2 days in the latter.


Unclassified, lactose-fermenting, urease-producing member of the family Enterobacteriaceae resembling Escherichia coli. Eight strains of an unclassified, oxidase-negative, lactose-fermenting, urease-producing, gram-negative bacillus were isolated from clinical material. With the exception of urease production, the biochemical characteristics of these organisms resembled those of Escherichia coli.


Effect of cetylpyridinium chloride on pathogenic fungi and Nocardia asteroides in sputum. The effect of cetylpyridinium chloride (CPC) on pathogenic fungi and Nocardia asteroides was studied. Sputa seeded with each of 11 organisms (Aspergillus flavus; Aspergillus fumigatus, Blastomyces dermatitidis, Candida albicans, Coccidioides immitis, Cryptococcus neoformans, Geotrichum candidum, Histoplasma capsulatum; Nocardia asteroides, Paracoccidioides brasiliensis, and Sporothrix schenckii) were treated with CPC and kept for 2, 5 and 9 days. The CPC reagent used (0.5% CPC and 0.5% sodium chloride) is one the Mycobacteriolgoy Branch at the Center for Disease Control added to sputa before shipping them to laboratories for recovery of mycobacteria. None of the organisms tested survived this treatment, and none was recovered on mycological or mycobacteriological media. Seeded sputa containing these organisms were also tested with a second CPC reagent (0.02% CPC and 0.5% sodium chloride) and held for 2, 5 and 9 days. A few colonies of A. flavus, A. fumigatus, and N. asteroides were recovered from these samples. Neither the morphology of the fungi nor their stainability by the fluorescent antibody method was affected by treatment with the reagent containing 0.5% CPC. However, the background material in smears from the 0.5% CPC-treated samples retained the conjugate, and this made weakly fluorescing organisms more difficult to detect. The 0.5% CPC treatment did not alter the morphology of N. asteroides or its ability to be stained with Kinyoun acid-fast stain.


Possibility of diagnosing meningitis by gas chromatography: cryptococcal meningitis. Cerebrospinal fluid (CSF) from eight patients with cryptococcal meningitis, from ten patients with viral meningitis, and from four control patients without meningitis were analyzed by electron-capture gas-liquid chromatography (EC-GLC). All cryptococcal specimens had similar EC-GLC profiles, and these differed from those of the controls. Viral EC-GLC patterns were different from those obtained with specimens from the patients with cryptococcal infection and from the controls. In addition, specimens from patients with various types of viral infections gave profiles that differed from each other. Two normal CSFs were inoculated with Cryptococcus neoformans; aliquots of these cultures showed an EC-GLC pattern very similar to that seen in CSF of patients with cryptococcal meningitis. The EC-GLC procedure is rapid, reproducible, and easy to perform and may hold promise as an additional aid in the diagnosis of cryptococcal infection.


Rapid recognition of group B streptococci by pigment production and counterimmunoelectrophoresis. Streptococci from clinical isolates were studied for their ability to produce pigment in stab cultures in Columbia agar. Serological grouping of these organisms was done by counterimmunoelectrophoresis using Burroughs-Wellcome antisera. In this group of isolates, 66 of the 68 organisms grouped as B by serological testing produced pigment in the Columbia agar stab cultures. Pigment was not produced by any of the other 36 streptococci studied (11 group A, 9 group C, 4 group D, and 12 nongroupable). The use of the Columbia agar stab culture is recommended as a rapid and simple test for recognition of group B streptococci. The counterimmunoelectrophoresis test is also suggested as a convenient, rapid, and sensitive method for grouping the streptococci.


Modification of the Minitek Miniaturized Differentiation System for characterization of anaerobic bacteria. The Minitek Miniaturized System (BBL) was modified for characterization of anaerobic bacteria. The modified system and the conventional Center for Disease Control method were used to test a variety of anaerobic bacteria, and results were compared. Tests performed by both techniques were indole and H2S production, esculin hydrolysis, nitrate reduction, and fermentation of glucose, mannitol, lactose, sucrose, maltose, salicin, glycerol, xylose, arabinose, mannose, rhamnose, and trehalose. The manufacturer's recommended procedure for the Minitek system was modified by using a new suspension medium (Lombard-Dowell broth) and an inoculum equivalent to the density of a McFarland no. 5 nephelometer standard. The Minitek results, recorded after 48 h, agreed satisfactorily with the conventional test results, usually recorded after 5 to 7 days of incubation. In the examination of 80 strains representing 22 different species or subspecies of anaerobic bacteria, with 16 biochemical tests performed in triplicate, 93.8% of the Minitek test results agreed with those of the corresponding conventional tests. Only tests for indole, H2S, and nitrate reduction gave less than 90% agreement. It was concluded that the modified Minitek system is a suitable substitute for the more expensive and time-consuming conventional procedure for determining carbohydrate fermentation and esculin hydrolysis by anaerobes. This system, when used in conjunction with other tests, can effectively aid in the definitive identification of commonly isolated anaerobes.


Rapid fermentation testing of anaerobic bacteria. Rapid tests for glucose, maltose, lactose, sucrose, and starch fermentation were performed on 112 strains of anaerobic bacteria. The tests were incubated under aerobic conditions,  and results were read within 4 h. An overall correlation of 89% was achieved between the rapid tests and the Virginia Polytechnic Institute method.


Acceleration of tetrazolium reduction by bacteria. Conditions were assessed which would permit more rapid recognition of bacterial growth than has been previously reported using tetrazolium salts. Microtitration trays were used. 2-(p-Iodophenyl)-3(p-nitrophenyl)-5-phenyltetrazolium chloride is rapidly reduced by respiring cells in tissue homogenates but is more toxic than other tetrazoliums when added to growing bacterial cultures. Phenazine methosulfate (PMS), an intermediate electron carrier, potentiates tetrazolium reduction. Growth was readily detected by the addition of these compounds after 3 to 4 h of incubation in Schaedler broth. The final concentration prior to addition to tray wells was 1.0 mg/ml for 2-(p-iodophenyl)-3(p-nitrophenyl)-5-phenyltetrazolium chloride and 0.06 mg/ml for PMS. Addition of 0.5 to 0.8 g of agar per liter of broth enhanced subsequent tetrazolium reduction.


Differentiation of Proteus mirabilis by bacteriophage typing and the Dienes reaction. A provisional typing schema based on sensitivity to 23 bacteriophages has been established for Proteus mirabilis. Seventy-three bacteriophages were isolated on strains of P. mirabilis (64), P. vulgaris (1), P. morganii (7), and P. rettgeri (1), but those isolated on P. mirabilis were the most useful in differentiating other strains of . mirabilis. From the 73 phages studied, the best 23 were chosen by computer analysis for the provisional system, which was then used to study P. mirabilis infections in a 500-bed general hospital. All patient isolates for 19 months were saved and then compared by bacteriophage typing and the Dienes reaction in a retrospective study. There was evidence for only three instances of cross-infection or -colonization during this time. Bacteriophage typing was very sensitive in differentiating strains, since 200 strains were differentiated into 113 different lysis patterns and 94% were typable. The Dienes reaction was useful at times but often gave reactions that were difficult to read or that changed when the tests were repeated. The bacteriophages described by Schmidt and Jeffries were also evaluated and proved useful in combination with ours. The value of bacteriophage typing was clearly established, and work toward a standardized schema for P. mirabilis should continue.


Clinical cholera caused by enterotoxigenic Escherichia coli. A woman returning from Mexico was hospitalized as an emergency patient with hypovolemic shock due to dirrheal disease of less than 1-day duration. Her clinical course was similar to that of severe cholera--she excreted greater than 60 liters of stool and urine in a 4-day period. The etiological agent was a non-enteropathogenic serotype but enterotoxigenic strain of Escherichia coli (063:NM). The patient responded both agglutinating and antitoxic antibodies against this strain and its enterotoxin. An "enteropathogenic serotype," 0111:B4, was also isolated but this finding had no etiological significance.


Significance of the isolation of Clostridium welchii from routine blood cultures. Clostridium welchii has been demonstrated in approximately 20% of contact plates taken from the antecubital fossa of 185 inpatients and outpatients and healthy staff. The highest incidence was in a group of 40 very ill patients. The isolation of the organism from blood cultures is not always of clinical significance. Skin preparation as at present practised is often inadequate to remove the spores when contamination is relatively heavy, for example, in bedridden patients.


Validation and standardization of Kveim test suspensions prepared from two human sarcoid spleens. Single lots of a Chase-Siltzbach type I Kveim test material from each of two sarcoid spleens and designated lot 5 of spleen K12 and lot 1 of spleen K13 have been validated alongside a single lot (lot 10) of a 'standard' suspension provided by Dr L. E. Siltzbach and prepared identically from the spleen of patient J (SPLEEN J) in New York. Additionally, a half-dilution of lot 5, K12, was included in this comparison. The reactivity of each suspension was assessed among patients with active and inactive sarcoidosis. The selectivity of each suspension for sarcoidosis was assessed similarly by comparison with results in patients with active and quiescent pulmonary parenchymal tuberculosis and in healthy subjects. All patients were closely matched and two Kveim tests were made in each subject according to a prearranged statistical design. The reactivity of the K12, K12 1/2 dilution, and K13 suspensions among patients with active and inactive sarcoidosis was closely similar to that with the 'standard' S10 suspension and in accordance with the expected proportions of reactions in patients at different stages of sarcoidosis. The K12, K13, and 'standard' S10 suspensions yielded a negligible proportion of positive reactions among patients with active and quiescent pulmonary tuberculosis and among healthy subjects: thus, as judged by these tests each suspension showed a high degree of selectivity for sarcoidosis. The results of this validation study are discussed in relation to the results of other studies in which lots 5 and 14 of K12 and early and late batches of a suspension prepared from another sarcoid spleen at the Commonwealth Serum Laboratories designated CSL and provided by Dr T.H. Hurley in Melbourne were employed. Using lot 5 of K12 positive reactions were found in an appreciable proportion of patients with Crohn's disease, ulcerative colitis, and tuberculous lymphadenitis. A closely similar rate of positive reactions was encountered among patients with Crohn's disease following tests with batch 0025 of CSL suspension and with another lot (lot 14) derived from spleen K12. A close concordance of results was obtained with lot 5(K12) and with batch 0042 CSL among patients with ulcerative colitis, but at a lower rate of reactivity. We conclude that positive reactions also occur in some diseases other than sarcoidosis and consider that the difficulties in determining the criteria for an acceptable test suspension become increasingly apparent as additional Kveim tests are made with one particular lot and with seqential lots of material from a 'validated' tissue source.


Localization of prolactin in chromophobe pituitary adenomas: study of human necropsy material by immunoperoxidase technique. In order to identify prolactin-producing tumours in human pituitary glands, 45 chromophobe adenomas, obtained from unselected necropsies, have been studied by various staining procedures including the immunoperoxidase technique for the demonstration of prolactin. The presence of immunoreactive prolactin was revealed in the cytoplasm of the tumour cells in six cases (13%), indicating that the occurrence of prolactin-producing adenomas is not rare. No correlations were established between tumours and clinical history. Two adenomas were detected in female and four in male patients. The age of the patients at necropsy ranged from 28 to 75 years. Three adenomas were associated with disseminated carcinoma, two with fatal liver disease, and one with diabetes mellitus, atherosclerosis, and pyelonephritis. Manifest endocrine symptoms were not disclosed, and endocrine investigations, including measurements of blood prolactin levels, were not undertaken. Thus, direct evidence is lacking as to whether or not these tumours were actively secreting prolactin. In the non-tumorous parts of the anterior lobes the number of prolactin cells was decreased in two cases, suggesting that prolactin released from the adenoma cells suppressed prolactin production in the non-tumorous pituitary. However, the number of prolactin cells of the non-tumorous adenohypophysis seemed to be unchanged in two and increased in another two cases. The present findings conclusively proved the existence of the prolactin-producing adenomas as a distinct entity. These tumours do not stain with acid or basic dyes, they are PAS or thionin negative, and do not contain immunoreactive growth hormone. Thus, by conventional staining procedures they are indistinguishable from other chromophobe adenoma types. Herlant's erythrosin and Brookes' carmoisine methods, claimed spedifically to stain prolactin cells, failed to provide reliable results, hence their use cannot be recommended in tumour identification. Immunoperoxidase staining of prolactin is the only technique which conclusively reveals the presence of immunoreactive prolactin in the cytoplasm of the tumour cells and permits diagnosis. It is proposed that this technique be introduced in pituitary morphological studies. Its application may lead to a better understanding of problems related to prolactin-producing tumours and their secretory activity.


A simple technique for observing germ tube formation in Candida albicans. Candida albicans is identified in most laboratories by the rapid formation of germ tubes when it is grown in human serum (Taschdjian et al, 1960). Demonstration of the production of germ tubes on solid media, 1% bactopeptone in 2% agar (Joshi and Gavin, 1974), and the appearance of other yeasts on a variety of carbohydrate media (Joshi et al, 1975) has led to the development of a simple technique for observing germ tube production.


Fine structure of synaptic vesicles in two types of nerve terminals in crayfish stretch receptor organs: influence of fixation methods. Crayfish stretch receptor organs were used. The standard procedure consisted of primary fixation with a 1.6% glutaraldehyde solution containing either buffer which was 60% hyposmotic or buffer which was isosmotic with the physiological solution (440 milliosmols), washing with isosmotic buffer, and postfixation with an isosmotic 1% osmium tetroxide solution. Under these conditions, we encountered two types of nerve terminals; small-vesicle terminals (SVTs) containing small elongated vesicles (300 approximately 330 A) and large-vesicle terminals (LVTs) containing larger round vesicles (430 A). Their location and physiological evidence suggest that SVTs and LVTs are inhibitory and excitatory, respectively. A hyperosmotic primary fixation solution due to increased glutaraldehyde concentration gave results similar to the standard procedures, while a hyperosmotic primary fixation solution due to increased buffer concentration caused shrinkage of the nerve terminal. A hyperosmotic buffer wash produced elongation of vesicles in SVTs and LVTs, while washing with hypoosmotic buffer rendered vesicles in both types round. Direct fixation with isosmotic osmium tetroxide yielded less elongated vesicles in SVTs and irregularly round vesicles in LVTs. However, under all conditions, vesicles in SVTs were smaller than those in LVTs. These results suggest that the consistent morphological difference in vesicles between SVTs and LVTs is size rather than shape, and it is important to standardize the osmolarity of primary fixation and washing solutions when discussing the differences of vesicle shape in various kinds of synapses.


Isolation of kappa-casein-like proteins from milks of various species. Kappa-Casein-like proteins were isolated from the milks of cow, goat, reindeer, horse, rat, and rabbit. When treated with rennin, all of the isolated kappa-casein components yielded para-kappa-casein-like bands on gel electrophoresis. The rate of cleavage of these components with rennin was determined by measuring material soluble in trichloroacetic acid (macropeptide). The curves were characteristic of a limited, specific attack by rennin on these proteins. The goat and reindeer kappa-caseins were nearly as bovine kappa-casein, but the cleavage of horse, rat, and rabbit kappa-casein-like components was much slower.


Enhanced fluoride uptake in enamel with a fluoride-impregnated prophylactic cup. A series of in vitro studies have been performed to study the release of tin and fluoride from an experimental prophylactic cup and the influence of this release on the fluoride content of enamel. It has been found that the release of fluoride from the prophy cup during a simulated prophylaxis results in a significant increase in the fluoride content of the treated enamel. It has also been observed that in those instances in which the fluoride-containing paste has a pH of less than about 3.8 this effect is partially additive to the increase in the fluoride content of enamel associated with the use of fluoride-containing prophylactic pastes. Collectively, these findings suggest that this prophylactic cup merits further investigation as a practical means of increasing the value of the prophylaxis phase of a multiple-fluoride treatment program for the partial control of dental caries.


Studies on hypoxia: XI. Long-term effects on the epiphyseal plate--a histomeric and radioautographic study. Rats were exposed to hypoxia for one week. The mean thickness of epiphyseal plates form control rats was 430 micrometers (mum) which was reduced to 313 mum in hypoxic rats. Radioautographic incorporation of 3H-phenylalaine by connective tissue cells in hypoxic rats was reduced up to 38% in control rats.