Combination radical surgery and multiple sequential chemotherapy for the treatment of advanced carcinoma of the testis (stage III). Nineteen patients with advanced testicular carcinomas (Stage III) were treated by a combination of multisequential chemotherapy with vinblastine, bleomycin, platinum, adriamycin, cytoxan, actinomycin D, and reductive surgery. Of the sixteen patients who were operated on 10 patients are alive and free of disease from 5 to 15 months. Two patients are alive with residual disease and four patients died. (Three were free of malignant tumor and one had residual malignant disease.) Three patients are still receiving preoperative chemotherapy. Pathology reported benign tumors in 8 out of 16 patients operated on. All these patients were previously treated with multiple chemotherapy. Several facts emerge from this study: (1) the great effectiveness of the combination of multisequential chemotherapy and reductive surgery to produce complete clinical remission in advanced testicular carcinomas (63.6%, 13 out of 19 patients); (2) the possibility of benign transformation of these tumors (50%, 8 out of 16); (3) the importance of the second-look surgery in testicular tumors.


The value of serial measurement of both human chorionic gonadotropin and alpha-fetoprotein for monitoring germinal cell tumors. Quantitative serial serum measurements of human chorionic gonadotropin (hCG) and alpha-fetoprotein (AFP) levels using sensitive double-antibody radioimmunoassays were performed in nine patients with germinal cell tumors before and during treatment. The sera of eight of the nine were found to have a hCG marker and five of the nine an AFP marker. The sera of four patients were found to have both. Serial serum levels of hCG, of AFP, or both were useful for monitoring disease activity during therapy in all nine patients. In two patients tumor masses failed to diminish during chemotherapy, but the tumor markers fell appropriately. At surgery one patient had a mature teratoma, the other a mature teratoma with a microscopic focus of an embryonal cell tumor. In one patient tumor reactivation was reflected by the emergence of only one of two previously elevated tumor markers. One patient had a rise in hCG, another a rise in both markers coincident with recurrence of tumor. Serial measurements of AFP and hCG are useful for following the response to therapy of germinal tumors, and can assist in making therapeutic decisions.


111Indium-bleomycin breast and axilla imaging. 111Indium-Bleomycin (111In-Blm), a new radiopharmaceutical, was administered intravenously to 37 patients with benign and malignant breast lesions. Early and delayed images of both the breasts and axillae were made, and results were correlated with physical examination, histopathology of the excised lesion, mammography, and thermography. In 18 patients with malignant disease, clinical examination of the breast and axilla correlated with histopathology in 78 and 54% of the cases, respectively. Images of the breast were accurate (true positives) in 83% of the cases. Images of the axilla were accurate in 62% of the cases. Mammography was correct and suggested malignancy in 88%, and thermography in 73% of the cases. In 19 patients with benign breast lesions, clinical examination of the breast and axilla correlated with histopathology in 68 and 95% of the cases, respectively. Scans of the breast and axilla were correct (true negative) 79 and 95% of the time, respectively. Mammography was correct, and suggested benignancy, in 53% and thermography in 25% of the cases. Imaging of the breasts using 111In-Blm appears to be as accurate as physical examination and mammography for palpable benign and malignant breast tumors. It is less accurate than mammography for microscopic malignancies. Axillary imaging does not appear to be worthwhile because many axillary metastases are too small for detection with current nuclear medicine instrumentation.


The radionuclide identification of tumors. Tumor-seeking radiopharmaceuticals have been employed in the diagnosis of primary neoplasms, in the detection of distant disease, particularly in the localization of tumor foci to facilitate biopsies and the planning of radiation portals, and in assessing the response to tumor therapy. At the present, there is no ideal tumor-scanning agent. However, several approaches appear to be useful and offer promise for further study. The greatest experience has been with Gallium-67, which has major utility in the staging of Hodgkin's disease, in the diagnosis of bronchogenic carcinoma, in the detection of certain metastatic brain tumors, in the identification of recurrent disease, and in the noninvasive diagnosis of leukemic complications. A number of radiolabeled antibiotic and chemotherapeutic agents have shown promise, including tetracycline and bleomycin. A major drawback, however, of these agents which is shared with Gallium-67 is that they appear to be sequestered by inflammatory as well as neoplastic tissue. A most intriguing approach is the use of radiolabeled antibodies to tumor-associated antigens. Animal and clinical experiments have employed antifibrin, antifibrinogen, anticarcinoembryonic antigen, and antiferritin. Theoretically, agents such as these should allow for greater tumor specificity.


Immunologic approaches to the diagnosis of cancer. A variety of immunologic approaches can be applied to the diagnosis of human cancer, and these have the potential advantages of specificity and sensitivity. Many tumor distinctive markers have been detected in the circulation of cancer patients. Cancer patients frequently have detectably depressed immune competence, but also have cell-mediated and humoral immune reactivity against tumor-associated antigens. Radiolabeling of specific antibodies may allow localization in vivo of tumors. With each of these potential immunodiagnostic approaches, it is necessary to systematically evaluate the assays and determine their value for specific clinical applications.


Enhanced incorporation of thymidine into DNA in the liver of intact and partially hepatectomized rats pretreated with 5-azacytidine. The administration of 5-azacytidine to rats resulted in enhancement of thymidine incorporation into liver DNA. Repeated doses of the drug caused a greater than 10-fold increase of thymidine incorporation. In 5-azacytidine-treated intact rats no changes in the activity of thymidine and thymidylate kinases were observed, whereas a marked depression of thymidine phosphorylase activity occurred. In intact animals the mitotic activity in the drug-treated livers changed slightly; however, 5-azacytidine administration before partial hepatectomy resulted in a dramatic increase of mitotic activity in 24-hr regenerating livers. In this case the observed increase in incorporation of thymidine was paralleled both by increased activity of thymidine and thymidylate kinases and by a decline in thymidine phosphorylase activity. The higher incorporation of radioactivity into liver DNA in intact rats pretreated with the drug can be accounted for, at least partially, by the lower cellular degradation of the injected radioactive thymidine used for labeling. In addition, alterations in the labeling and in the mitotic activity of hepatocytes and nonparenchymal cells caused by 5-azacytidine should be taken into consideration.


Experimental infection of a cat kidney cell line with the mouse mammary tumor virus. A cat kidney cell line, CRFK-F2, was successfully inoculated in suspension and in monolayer culture with a purified mouse mammary tumor virus derived from RIII milk. The virus produced by the infected cells was identified by immunogluorescence, electron microscopy, and RNA-directed DNA polymerase assays; it was a B-type virion that did not cross-react with mouse or feline leukemia-sarcoma viruses, had spikes on its envelope, and had a RNA-directed DNA polymerase reaction that was typical of mouse mammary tumor virus. The producing cells were identified as cat cells by chromosome number, cytotoxic assays, and isoenzyme migratory patterns. A standardized method for the in vitro inoculation of cat cells is described that presently permits highly reproducible results. For the first time, the mouse mammary tumor virus is seen replicating in cells from another species, thus offering an opportunity to study the kinetics of infection of that virus.


Retinal alterations induced by continuous light in immature rats. II. Zinc iodide-osmium tetroxide (ZIO) reactive sites in the rod outer segments. Immature albino rats were subjected to (a) continuous illumination for 5-9 days, or (b) continuous illumination followed by prolonged darkness. Their electroretinographic responses and the ultrastructural characteristics of the rod outer segments, as revealed by a mixture of zinc iodine-osmium tetroxide (ZIO) at different temperatures, were studied and compared with those of a control group maintained in a cyclic rhythm of light and darkness. Noteworthy differences in the distribution of ZIO reactive sites were observed in the rats exposed for 5-9 days to continuous illumination (no electroretinographic responses) as compared with normal controls. At 4 degrees C, ZIO staining was negative in the rods of illuminated rats whereas at 20 and 60 degrees C it was positive inside the tubular and vesicular structures. After prolonged darkness, in rats with a partial electroretinographic recovery and ultrastructural restoration, the ZIO reaction showed a similar pattern to that observed in the control group, ZIO deposits being found both in the intra- and extradiscal spaces.


[Serum protein differences between ambulatory and bed-ridden subjects]. Total proteins as well as 7 individual protein components were evaluated in the blood serum of 19 subjects, first ambulatory, then recumbent. A concentration decrease of 7% has been found for the latter, independently of the nature of the various proteins studied.


[Induction, by 5-bromo-2'-deoxyuridine, of a "foamy" virus previously undetected in hamster cells transformed by SV40]. TSV5 clone 2 cells in normal conditions of culture contain only an expressed RNA virus (R-type virus). However, exposure of the cells to 5-bromo-2'-deoxyuridine with dexamethasone, induced synthesis of a syncitium-forming ("Foamy") virus. In other hamster cell lines, the same treatment fails to induce a "foamy" virus. The origin of this "foamy" virus is discussed.


Electron microscopic immunohistochemical identification of endothelial cells in the rabbit. Antibody to tissue factor apoprotein was adsorbed against gamma-globulin and coupled to horseradish peroxidase; this complex was applied to various rabbit tissues. The distribution of the peroxidase marker then was observed by electron microscopy. We examined fixed or frozen sections, as well as breis of aorta, vena cava, brain, heart, lung, liver, spleen, kidney, bone marrow, mesothelial gut lining, erythrocytes, and platelets. All endothelial cells that had been exposed to the antibody complex were positive and in all cases only the endothelial cells showed localization of the electron-dense reaction product. Tissues that had been incubated with complexes prepared with gamma-globulin from animals not immunized with tissue factor apoprotein showed no staining. Prior treatment of the tissue with uncoupled anti-tissue factor gamma-globulin blocked binding by the coupled antibody. In blood vessel preparations that had been specifically designed to expose the media to the anti-tissue factor complex, medial smooth muscle cells and connective tissue showed no reaction product. Parenchymal cells of the other other organs mentioned likewise were devoid of reaction product. Similarly, the leukocytes and platelets occasionally observed in vessel lumens showed no evidence of binding. Platelets adhering to arterial subendothelial structure after injury also were unreactive. These findings suggest that in normal rabbits anti-tissue factor-horseradish peroxidase complex combines selectively with endothelial cells.


Coagulation of serum by glutaraldehyde.

Coagulation of serum by glutaraldehyde. A semiquantitative method for the determination of the gamma-globulin in human serum was developed based on the observation that dilute glutaraldehyde solutions at serum pH first attack the most basic serum proteins, leading to a polymer (clot) consisting mainly of gamma-globulins. The clotting ability of serum by glutaraldehyde thus caould be used as an indicator of the gamma-globulin concentration.


Double-antibody enzyme immunoassay applied to human alpha 1-fetoprotein. We report the development of a double-antibody system for enzyme immunoassay of human alpha 1-fetoprotein. Equilibrium and sequential saturation procedures were evaluated and compared with a similar double-antibody radioimmunoassay. With our method, 3 mug of alpha 1-fetoprotein per liter can be detected. The dose-response curve covers a 100-fold range of analyte concentrations. Within-run and between-run coefficients of variation are respectively 6.9% and 10.0%. Analytical recovery of added antigen is 99.5 +/- 10.6%. Results are routinely obtained in 24 h but can be obtained in less than 8 h. Alpha 1-fetoprotein was measured in a variety of clinical samples, including sera from 488 pregnant women. The advantages of the present method over radioimmunoassay and other enzyme immunoassay procedures are discussed.


[Variations of the concentrations of certain glycoproteins in maternal serum, fetal serum and amniotic fluid during pregnancy (author's transl)]. The authors investigated systematically the variations during normal pregnancies of the concentrations of alpha-1-antitrypsin, orosomucoid, transferrin and alpha-fetoprotein simultaneously in maternal serum, fetal serum and amniotic fluid. The role of certain factors such as the gestational age birth weight, placental weight and pairty were studied with regard to variations in the concentrations of each of these proteins. This research permitted the definition during pregnancy of the normal concentrations for these four proteins and allowed us to learn more about protein exchanges between fetal blood, maternal blood and amniotic fluid. There exists a difference between the concentrations of alpha-1-antitrypsin and of orosomucoid found for primigravidae and for multigravidae. The role of these glycoproteins in preventing the mother from rejecting the fetus (insofar as the fetus may be considered as an allograft) is discussed.


[Identification of proteins by immunochemical methods after electrophoretic runs on different supports (author's transl)]. The determination of the location of serum proteins after electrophoresis on various supports and after immunoelectrophoresis has been achieved by immunochemical methods with specific antisera. The present study describes the technique, gives details of various identified fractions and presents some figures. The method allows us to study the constitution of the "beta-gamma block" observed on paper and acetate supports after electrophoresis of cirrhotic sera and to superimpose the patterns of apparently distinct fractions on polyacrylamide gel.


The association of alpha2-macroglobulin with lymphocyte membranes in chronic lymphocytic leukaemia and other disorders. The association of alpha2-macroglobulin (alpha2M) with the surface membranes of human peripheral blood lymphocyte preparations has been investigated by the direct immunofluorescent technique. The percentage (and total number) of lymphocytes with detectable alpha2M on their surface is significantly increased in chronic lymphocytic leukaemia. The incidence of alpha2M-positive cells in normal and pathological conditions closely parallels that observed with conjugated antiserum to the kamma and gamma light chains of human immunoglobulin. It bears no relationship however to the plasma alpha2M levels or to the age of the donor. Additional blocking studies with aggregated human IgG, soluble antigen-antibody complexes and the F(ab')2 moiety of the anti-alpha2M, and indirect immunofluorescent studies with the latter, indicate that the fluorescence observed is not due to interaction of the conjugated reagent with Fc or C3 receptors.


Potentiation of IgE-mediated cutaneous reactivity and blood leucocyte histamine release by deuterium oxide. A previous study (Gillespie & Lichtenstein, 1972) demonstrated that there was potentiation of histamine release from human peripheral blood leucocytes following exposure to antigin or anti-IgE in deuterium oxide (D2O). The current study confirms the results with human leucocytes and indicates that the degree of histamine release due to anti-IgE or its potentiation by D2O appeared independent of the serum IgE concentration of the cell donor. Further studies demonstrated that the peripheral blood leucocytes from monkeys with a sufficient degree of IgE-mediated reactivity to Ascaris antigen-released histamine following exposure to that antigen. This leucocyte histamine release occurs in animals with immediate-type cutaneous and respiratory reactivity following challenge with this antigen. Peripheral blood leucocytes from certain monkeys release histamine following exposure to anti-human IgE. Both of these Rhesus leucocyte responses are potentiated by D2O. This potentiation of histamine release in vitro in two species by D2O was compared with the potentiation of cutaneous reactivity in IgE-mediated cutaneous reactions in two species. The addition of D2O to Ascaris antigen or anti-IgE increased the end-point cutaneous titres to these stimuli in Rhesus monkeys and the addition of D2O to Ascaris or ragweed antigen increased the end point cutaneous titre to these reactants in allergic dogs. D2O did not potentiate cutaneous reactivity to histamine in either dog or Rhesus monkey.


Characterization of a new antigen-antibody system associated with hepatitis B. The occurrence of a new antigen-antibody system was recently described in hepatitis B surface antigen (HBsAg) positive sera with different specificities, one of which was designated e. These specificities are related as demonstrated by the presently utilized e reagents not discriminating between them in immunodiffusion tests. Hence, they are collectively referred to as e in this report. The occurrence of e antibodies in sera containing antibodies against hepatitis B surface antigen (anti-HBs) is reported. The e antibodies were found to move as 7S globulin on rate zonal centrifugation. The e precipitate was demonstrated on the cathode side of the HBsAg precipitate on immunoelectrophoresis consistent with e antigen migrating in the gamma-globulin region. The e antigen is reported to have an S value of 11-6 +/- 0-6 (s.d.) and a buoyant density of 1-291 +/- 0-003 (s.d.), hence differing from HBsAg in all respects studied thus far.


Species-specific tissue antigens. III. Immunological relationships of enzymic antigens in various species. Immunological relationships have been investigated with acid and alkaline phosphatases, cystine aminopeptidase, beta-acetylglucosaminidase, beta-glucuronidase, catalase and L-glutamate dehydrogenase of human, monkey, mouse, rat, rabbit, dog, cattle, sheep, cat, pig, guinea-pig and chicken organ extracts by means of immunodiffusion and immunoelectrophoresis. Extensive cross-reactions among the antigens of most of the enzymes were observed. However, enzymic proteins of acid and alkaline phosphatases, cystine aminopeptidase, beta-acetylglucosaminidase and beta-glucuronidase were found to possess primate and/or human-specific antigenic determinants.


The effect of anti-alpha2-macroglobulin on K-cell cytolysis and T- and B-cell rosette formation. The effect of antisera to human alpha2-macroglobulin (alpha2M) on the ability of human peripheral blood lymphoid cells to lyse antibody coated target cells (exert a K-cell effect) and form T(E) and B(EAC) rosettes has been investigated. Pretreatment of lymphoid cells with these antisera inhibited their K-cell activity but had no effect on their capacity to form T or B rosettes. The inhibitory activity of the anti-alpha2M sera was always reduced following absorption on a CnBr column to which human alpha2M had been coupled. The absorbed antibody, recovered by acid elution reacted strongly with human alpha2M in gel diffusion precipitin analysis and caused a highly significant inhibition of K-cell cytotoxicity. It was further shown that the inhibitory activity of the anti-alpha2M was localized exclusively in the IgG fraction and was greatly reduced following pepsin digestion.


Cell-mediated immune response in mice treated with steroidal contraceptives. The splenomegaly assay (Simonsen, 1962) was standardized using different strains of rats and mice. Male Wistar rat (donor)-female Swiss mouse (host) was found to be the suitable combination that could be employed in subsequent experiments to study the potential of contraceptive steroids to alter CMIR. The index of splenomegaly appeared to increase in case of mice treated with combination oral contraceptives (Ovulen, Ovral or Enovid). The differences observed, however, neared significance only in the case of Ovral (0-05 less than P less than 0-1). Neither chlormadinone acetate nor megestrol acetate significantly altered the index of splenomegaly.


Studies with the macrophage migration inhibition (MMI) test in patients with malignant disease. Macrophage migration inhibition (MMI) tests have been carried out using lymphocytes from twenty-six patients with malignant disease and from twenty-two other subjects. The antigens used were the same as those employed in the macrophage electrophoretic mobility (MEM) test. The results indicate that the inforation obtained using the MMI test is similar to that found with the MEM test.


The fluorescence localization of mouse satellite DNA in interphase nuclei. Nuclei from various mouse tissues exhibit a pattern of fluorescence characteristic of the cell type when stained with the fluorescent compound Hoechst 33258. When such preparations are hybridized in situ with 3H-RNA complementary to the A-T rich satellite of mouse, it is clearly seen that only the fluorescent regions of the nuclei contain the satellite DNA. Thus Hoechst 33258 allows the precise localization of satellite DNA at all stages of the mouse cell cycle.


Method for the determination of mean densitometric profiles of chromosomes: application to human chromosomes stained by quinacrine mustard, ethidium bromide or by the Feulgen reaction. When comparing the densitometric profiles of corresponding chromosomes registered from different metaphases or homologous pairs, one is always faced with the variability of their length and overall height. This makes difficult the quantitative comparison of a given chromosome treated by various staining procedures.--A simple and rapid method has been developed for normalizing the densitometric profiles and averaging them in order to obtain a "mean density pattern" of each chromosome. The analysis involves: photographic images, digitalization of the densitometric profiles and processing of the data by a mini-computer.--The method, based on a linear relationship between the area of the densitometric profiles and their length, has been applied to five human chromosomes (1, 2, 6, 12 and 16) stained by ethidium bromide, quinacrine mustard (with or without acidic hydrolysis), pararosaniline and bisaminophenyl-oxadiazole (Feulgen reaction).


[Localization and significance of the Q bands observed on mitotic chromosomes of the amphibian Pleurodeles waltlii Michah. after coloration by quinacrine mustare (author's transl)]. Metaphase plates from tailbuds of Pleurodeles waltlii embryos (stage 34) with or without preceding cold treatment were obtained by squashing followed by quinacrine mustard staining. In both cases, caryograms were established and sites of Q bands located.--Most of the secondary constrictions exhibit very high fluorescence. In general, it is the same for most centromeric parts, but their fluorescence is quenched very strongly by cold treatment.--The proximal part of the long arm of chromosome VII and satellites of chromosomes III and XI exhibit dull fluorescence. All these sites are compared with heterochromatin localization. The relation between banding and base composition or non-histone proteins interactions is discussed.


[Effect of prajmalium bitartrate and procaine amide on ventricular extrasystoles (author's transl)]. Prajmalium bitartrate (Neo-Gilurytmal) was given to 19 patients, who had either runs of ventricular extrasystoles or frequent extrasystoles. The E. C. G. was continuously monitored on magnetic tape. At a dosage of 80 mg/d (20 mg every  six hours) a significant reduction in ventricular extrasystoles was demonstrated on 20 separate occasions. After two hours extrasystoles decreased to 63% of the initial level. After six hours practically the full effect of the drug had been reached; the reduction in extrasystoles stabilized after 12 hours at an average of 30% of initial level. Ten hours after the last tablet had been taken the frequency of extrasystoles had risen to 57% of initial level. Comparison with procaine amide (3 g/d) in 14 patients revealed no significant difference between the two drugs. Prajmalium bitartrate was well tolerated, while three patients receiving procaine amide complained of nausea and gastrointestinal symptoms, so that treatment had to be discontinued in two. In one patient, receiving procaine amide by continuous drip, there were reversible joint pains, resembling lupus erythematodes.


Kindling in the frog: development of spontaneous epileptiform activity. The present report is the first demonstration of the kindling phenomenon in a non-mammalian, poikilothermic species. Brief (2 sec) localized electrical stimulation was delivered to the hippocampal cortex of one hemisphere of the partially paralyzed bullfrog, Rana catesbeiana, at current levels just sufficient to elicit brief after-discharge (AD) on first application. Succeeding stimulations, once per hour, gave rise to more and more prolonged AD involving first the side stimulated (1 degree focus) and secondly the opposite hemisphere (2 degree focus). In addition to these stimulus-dependent alterations, after several hours spontaneous, paroxysmal discharges occurred in  interstimulus intervals. They appeared first in the stimulated hemisphere, then as evoked spikes in the secondary hemisphere, and finally independently on the two sides. The waveshape of the spontaneous epileptiform potentials exhibited an extraordinary constancy from complex to complex which was specific to each animal. Such wave-shape coherence implies a constancy to both the anatomical substrate and the sequency of synaptic engagement each time spontaneous paroxysmal activity invades the network. The model is especially suitable for analysis of temperature-dependent biochemical events responsible for this functional transformation.


A new technique for long-term recording of eye movements in infants. A device for recording eye movements from infants over long time periods is described. The device consists of an infrared light-emitting diode and a phototransistor mounted in a small package which covers one eye. The eyepiece is insensitive to EEG, EKG, or EMG potentials, and avoids irritation of the infant's skin from application of electrode paste. Detection of eye movements compares favorably to conventional eye movement electrodes.


Anatomical organization of the phasic activity produced by reserpine at the level of the oculomotor system. The organization of the pathways responsible for the transmission of phasic electrical activity at the level of the oculomotor system was studied in the encéphale isolé cat which was injected with reserpine or exhibited spontaneous phases of paradoxical sleep. At the level of the VIth nuclei there are both ipsilateral and contralateral connections deriving from each generator. The ipsilateral pathway transmits an "inhibiting" potential eliciting electromyographic inhibition of the ipsilateral rectus muscle, and the contralateral pathway, an "activating" potential eliciting activation of the corresponding lateral rectus muscle. A medial saggital section from frontal plane APO, extending caudally to the VIth nuclei at frontal plan P9 is necessary to suppress the bilateral synchronization of phasic activities recorded from the VIth nuclei and the activation of the lateral rectus muscles. The areas responsible for phasic activity recorded at the level of the central visual and oculomotor systems have been delimited through brain transections. The pathways responsible for the transmission of phasic activity at the level of the IIIrd and IVth nuclei are contiguous with the ponto-geniculate pathways. This was demonstrated by electrocoagulation.


The human frequency-following response: its behavior during continuous tone and tone burst stimulation. The frequency following response (FFR) in humans has been elicited by continuous tones as well as tone bursts. The responses were observable over a range of frequencies extending from 70 c/sec to greater than 1.5 kc/sec. The threshold for the continuous response has a mean of about 40 dB SL and is perhaps several dB lower for the burst response. The response threshold as a function of sound pressure level (SPL) increases rapidly at frequencies below 125 c/sec and above 1 kc/sec. These thresholds are particularly sharp at the lower frequencies. Narrow-band noise that just produces subjective tone masking also masks the FFR. The latency of the FFR is about 6 msec when the tone burst intensity is 30 dB over FFR threshold. Comparison with click response latencies indicates that the onset of the FFR corresponds with early waves IV and V.


The scalp topography of potentials associated with missing visual or auditory stimuli. Averaged potentials time-locked to regularly presented visual and auditory stimuli and to the occasional random deletion of a stimulus were recorded from a scalp electrode array overlying the left hemicranium. The major components of visual and auditory evoked potentials and of the potentials associated with missing stimuli (MSP) were measured and their amplitude distributions depicted in the form of isopotential maps. The N1 components of the VEP and AEP had distributions compatible with sources in and near the respective primary cortical projection areas. The P2 components were more widely distributed and could be attributed in part to generators within modality specific association areas. The MSP comprised two main components, an initial negativity (NMSP) and a later positive wave (PMSP). The NMSP distributions were different in the visual and auditory modalities, and were similar to the respective EP topographies. The NMSP appeared to reflect a more powerful contribution of association areas than did the evoked responses. The PMSP topography was modality unspecific with distributions which were maximal over the parietal region. The possible functional significance of the NMSP and PMSP was considered in the light of their timing and topography.


Maturation of the somatosensory evoked potentials in normal infants and children, with special reference to the early N1 component. The early components of the somatosensory evoked potential (SEP) were analysed in 39 normal newborns in REM sleep, 35 normal awake children between 1 month and 9 years and 16 normal awake adults. Electrical pulses were delivered at random intervals to fingers of the contralateral hand. The SEP were averaged from several electrodes in the parietal scalp focus and several runs were compared to estimate precise latencies and durations (Fig. 5, 6, 7). The system bandpass extended to 3 kc and many precautions were taken to exclude interference. The limb temperature was carefully maintained at normal value in order to avoid undue slowing of peripheral conduction velocity. The SEP presented markedly increased latencies for near-threshold stimuli (Fig. 3) while becoming even more focalised (Fig. 1). Background data were obtained in order to standardize the parameters for the maturation study (Fig. 2, 3, 4). In the children, the early negative component was found to undergo progressive changes. It was only at the age of about 8 years that the adult pattern was approached (Fig. 8, 9, 10, 11). The duration of the early negative N1 component decreased quite progressively. The latencies to onset and to peak were also shown to follow a highly consistent pattern when the body length of the subjects was taken into account (Fig. 9, 10). By dividing the data by the body length, functions were obtained which could be said to apply to a "standard" individual whose body length did not change from birth to adulthood and remained at one meter. Such plots made it possible to appreciate the true extent of the SEP maturational changes from birth. The quantitative data thus provided should serve as a useful reference for subsequent studies on developmental changes of the brain and for clinical applications to diseased children.


The human magnetoencephalogram: some EEG and related correlations. Simultaneous magnetoencephalographic (MEG) and electroencephalographic (EEG) data were recorded from six normal adult subjects. MEG signal strength and EEG voltage level appear to be linearly correlated. Spectral analysis suggested that the MEG and EEG data were produced by similar but non-identical generator systems. A vertex region magnetic averaged evoked response to flash was recorded in one (of four) subjects, consisting of a waveform similar to but out of phase with the simultaneous EEG averaged evoked response, such that cortical negativity was correlated with a magnetic field directed into the scalp. Eye movement artifact, which can seriously compromise EEG recordings, does not appear to be a major problem in MEG recordings.


Effect of flurazepam on sleep spindles and K-complexes. In this research, a quantitative study of the EEG from 5 subjects permitted a detailed analysis of the effect of 30 mg of flurazepam administered over 7 nights. Four placebo baseline nights and 3 placebo withdrawal nights were also recorded. For 4 of the subjects, a nondrug and nonplacebo follow-up record was obtained 4 to 6 weeks later. The subjects were 4 females, 1 male, age range 23-42. All complained of either sleep onset greater than 45 min, sleep length of less than 6 h, or two or more sleep awakenings. Compressed spectral analysis yielded a computer-generated somnogram on each of the 15 nights of sleep, and an automated spindle detector was used to count and measure the duration of spindle bursts with frequencies of 12.25-15.5 c/sec on baseline nights 3 and 4, drug nights 1, 2, 3 and 7, on the 3rd withdrawal night, and on the 4-6 week followup record. K-complexes were scored visually on the 4th baseline and 7th drug nights. There were no significant differences in spindle rate per minute among baseline nights and the follow-up record. By the 2nd drug night, spindle rate had significantly increased over the baseline rate. Linear contrast analysis indicated there was a significant increase of spindle rate over drug nights. All 5 subjects showed this pattern of increase. In contrast to the increase in spindle activity, the rate per minute of K-complexes significantly decreased during drug administration.


Changes in amplitude of the EEG induced by a photic stimulus. Occipital EEG responses to a single photic flash were studied by using a complex demodulation technique. Individual EEGs were first digitally filtered by making use of the Ormsby band-pass filter. Envelopes of the alpha (8-12 c/sec) and of the low frequency (3-7 c/sec) activities were obtained in terms of the demodulation calculation. Both individual and averaged data of raw, filtered, and demodulated EEGs were utilized for the present study. Two typical kinds of EEG responses were observed. One showed clearly the blocking as well as the enhancement of the alpha activity. The alpha activity first exhibited a reduction in amplitude in the range 50-200 msec after the stimulus (the minimum amplitude at about 150 msec). Then an evocation, rather than merely a recovery, of the alpha was observed in the range 500-800 msec (maximum amplitude at about 650 msec). The other typical example showed the low frequency evoked potential prominently. This response appeared almost locked to the stimulus and showed roughly a bi-phasic pattern in the time range where the alpha blocking took place (30-200 msec). Phases of the evoked low frequency component were almost locked to the stimulus but no distinct phase coherence was observed for the enhanced alpha wavetrains. Detection by demodulation of the phase change of the alpha was critical for a single EEG. It is pointed out that changes of the alpha must be studied on individual EEGs.


Biological signal detection by the autocorrelogram and a recurrence frequency method. The signal detection property of the autocorrelation function (ACF) is conventionally derived on the assumption of two conditions that may not be present in biological time sequences: (1) the measured variable is assumed to be the sum of a noise and a signal function; (2) the signal function is assumed to continue throughout the sequence. In neuroelectric sequences of all-or-none responses the first assumption would not be appropriate if most of the elements seen by the recording electrode were accessible to the signal input. The sensitivity of the ACF was investigated in simulated sequences in which the random noise was entirely replaced by a periodic square wave signal function for a small number of repetitions and the results compared with a recurrence frequency function (RFF) developed during this work; under these conditions the ACF proved to be significantly inferior. Sensitivity of the ACF was the same for a pure signal function as for an additive function of signal and noise but sensitivity of the RFF was severely degraded for the latter. A variety of neuroelectric sequences was studied and many were found in which strong rhythmic processes revealed by RFF were not detected by the ACF. It is suggested that both methods be used when the signal function may be present during only a small part of the data-collection period and when it is not known to what degree the measured variable may be the sum of a noise and signal function. When an additive function is known to exist, the ACF will be the method of choice; for pure signal functions REF will be more sensitive.


Correlation between the dischanges of two simultaneously recorded motor units and physiological tremor. Simultaneous recording was performed from pairs of motor units either in a small intrinsic hand muscle (first dorsal interosseus, 6 subjects, 37 records) or in two synergistic calf muscles (gastrocnemius and soleus, 6 subjects, 50 records). The two motor units were recorded by means of two electrodes inserted into the muscle during stationary isometric contractions of different strengths. Cross-correlograms were calculated in two ways: (1) between spike trains of two motor units; (2) between either spike train and the peaks of the tremor record. The cross-correlation between pairs of motor units showed a consistent tendency to synchronization in the hand as well as in the calf muscles. This synchronization was higher than could be expected from the random activity of asynchronously firing motor units. The amount of synchronization was correlated with the amplitude of physiological tremor: the stronger the tremor, the higher the synchronization coefficient. The cross-correlation between a spike train and the peaks of the tremor records showed a higher probability of unit firing 30-60 msec prior to the tremor beats. It is concluded that the tremor force produced by one muscle or a pair of synergistic muscles is the result of the synchronized activity of motor units. This hypothesis is supported by some characteristic changes in the amplitudes and rates of tremor in Parkinson's disease and myopathies. In these diseases the pathological changes in tremor rates simply reflect underlying changes of the motoneuronal firing rates.


EEG changes during spontaneous and controlled menstrual cycles and their correlation with psychological performance. 1. EEG (occipito-central, anterior and posterior temporo-central), blood concentration of hormones (oestradiol, progesterone, prolactin, folliculo-stimulating (FSH) and luteinizing (LH) hormones) and psychometric tests (reaction time, concentration, simple calculations, visual orientation and simple motor performance tasks) were recorded from 16 young women during a spontaneous menstrual cycle (s.c.w. group) and from 16 women taking combined oral contraceptives (o.c.w. group). All data were collected every second day except Sundays. 2. The EEGs underwent power spectrum analysis. Power and weighted mean frequency were determined for different frequency bands (theta, alpha and beta). The most constant finding in the s.c.w. group was a slight increase of mean alpha frequency during the luteal phase until menstruation (mean of occipital alpha acceleration = +0.3c/sec). The average time course of this acceleration fitted the time course of progesterone increase, but no direct correlation could be established. No such acceleration of alpha frequency could be found in the o.c.w. group, but the mean frequency in this group was about 0.5c/secslower than that of the s.c.w. group. In addition, minor cyclic changes of EEG paramenters in the s.c.w. group were observed in the occipital and temporal theta activity, the temporal alpha activity, but none in the beta range. 3. Reaction times to simple tasks, simple arithmetic, etc. were slightly but significantly decreased in the s.c.w. group during the luteal phase with a minimum on the day before or during menstruation. No such changes were observed in the o.c.w. group, which performed, in all tasks, slightly slower and slightly less correctly than the s.c.w. group. 4. The individual relative accelerations of alpha frequency and shortening of reaction times were positively correlated with each other.


The frequency-following response in subjects with profound unilateral hearing loss. Frequency-following responses (FFRs) evoked by 500 c/sec tone bursts 14 msec in duration, presented at 50 dB SL were recorded from Cz--A1 and Cz--A2 electrode derivations using eight subjects with normal bilateral hearing, and eight subjects with profound unilateral hearing loss. Monaural stimulation of either ear in normal subjects, and of the unimpaired ear in hearing-loss subjects, evoked larger responses from the ipsilateral electrode derivation than from the contralateral electrode derivation. Stimulation of the impaired ear in hearing-loss subjects evoked no response. Binaural stimulation in normal-hearing subjects evoked Cz--A1 and Cz--A2 responses of equal magnitude, each larger than either the ipsilateral or contralateral monaural response, but each slightly smaller than the sum of the ipsilateral and contralateral monaural responses. Binaural stimulation in hearing-loss subjects evoked responses equivalent to those obtained monaurally. The results provide evidence of binaural interaction in normal-hearing subjects and indicate that FFR arises from at least two separate symmetric neural sources, possibly by iterative activation of brainstem evoked response (BER) generators.


Study of contingent negative variation and the post-imperative wave in the presence of interference. The aim of this investigation was to study ways of inducing prolonged contingent negative variations (CNV) in human subjects. Fourteen subjects underwent the following experimental procedure: 1st day: 2 CNVs in control conditions were recorded (16 successive trials, each with standard S1--S2 paradigm). 2nd day: 5 successive CNVs were recorded:(1) control; (2) labyrinthine stimulation before 1st, 5th, 9th and 13th trials with simple mental calculation performed between S1 and S2, i.e. during CNV; (3) mental calculation only; (4) labyrinthine stimulation only; (5) control. None of the three interference situations caused any significant changes in the mean and maximal CNV amplitudes measured between S1 and S2. On the other hand, the post-imperative part of the CNV was significantly (P less than 0.025) prolonged but only in the situation where 2 types of interference were applied at the same time. This increase disappeared with removal of the double interference. This change, which matches that observed spontaneously and reversibly in certain mental patients in the acute phase of their illness, seems to reflect a transient alteration of the psychophysiological state of the subject. The possibility of inducing these changes in a normal subject could serve as a means of determining the individual threshold of stress.


Spatio-temporal organization of EEG in premature infants and full-term new-borns. Inter- and intrahemispheric EEG relationships were studied as a function of maturation in new-born infants. The spatio-temporal organization of EEG activity obtained during the two stages of sleep described in new-born infants - quiet sleep and active sleep - was analysed in 9 full-term new-born infants and 5 prematures (6 records). EEGs were recorded through transverse parietal montage made up of six bipolar derivations. Two epochs of 1.5 min each - successively obtained in both stages of sleep - were digitized, filtered in three frequency bands (beta, theta and delta) and computer-processed according to two methods, factor analysis and rhythms averaging. 1. The following EEG characteristics were found in both groups: (a) Instability of frequency within each frequency band (beta, theta and delta). However, theta activity was the most stable of the three, especially in quiet sleep. (b) Variability of topographical organization (i.e., localization of maxima of potentials) from one moment to another in both stages of sleep and in the three frequency bands. In spite of this intra-individual variability some sort of "average structure" was found in all infants; it was characterized by usually lateral and often symmetrical positions of the maxima of potential on the two hemispheres. For the same infant this structure was the same in the three frequency bands. (c) Poor interhemispheric relationships under all conditions, as well as poor interregional links in one hemisphere. 2. Some EEG characteristics differentiated the two groups and thus seemed to be related to maturation. Compared to full-term newborns the premature group showed: better stability for all three activities, especially for beta activity; higher frequency and larger amplitude of beta activity; better inter- and intrahemispheric relationships; fewer differences related to sleep stages. These results are discussed in terms of organization of the underlying cortical generators. The authors suggest that the active areas would increase in number and in surface with maturation, whereas the links between these different separate areas would remain very poor in the human full-term new-borns as well on one hemisphere as across hemispheres. This last finding would argue against the hypothesis that the corpus callosum which, as is well known, matures early, plays an important role in the establishment of interhemispheric links.


Detection of cyclic sleep phenomena using instantaneous heart rate. The development of the Heart Beat Domain and the Fourier transform of the Heart Beat Domain (which we call the Beatquency Domain) has provided new and useful tools for the quantitative analysis of sleep level patterns. This method of analysis has produced remarkable intersubject as well as intra-subject consistency and the only physiologic parameter required in the analysis is beat-by-beat heart rate. This analytical tool was designed to aid in the detection of sleep cycles, or more specifically, the rhythmic transitions from REM+ (awake Stages 1 and REM combined) to NREM (Stage 2, 3 and 4 combined) over a normal night of sleep. Employing this method on minute-by minute sleep recordings from 9 normal sleep subjects, 2 complete nights each, we were able to distinguish between the REM+ and NREM stages with an average accuracy of approximately 80%. Considering that beat-by-beat heart rate was our only criteria, we felt that the algorithm performed with significant success.


Time of day effects with standardized exercise upon subsequent sleep. The effect standardized exercise had upon sleep was studied with eight subjects. A pilot study assessed individual work capacity by the sub-maximal estimation of VO2 max. In the main study each subject performed the exercise, once a.m. and once p. m., on different days. Sleep was scored into stages and an additional sub-division of stage 2 containing 10-20% by time of delta. Comparison with baseline showed no significant whole night changes with any criteria following either a.m. or p. m. exercise. After p.m. exercise there was a significant increase in stage 3 for the first half of the night. It was concluded that ensuing wakefulness following early daytime exercise is sufficient for recovery, but late daytime exercise may result in an intrusion of recovery into initial sleep. Sleep is not seen to be necessary for recovery from muscular fatigue.


The thyroxine-binding properties of rat and rabbit serum proteins. The thyroxine (T4) binding properties of rat and rabbit serum proteins were studied using a gel equilibration technique, gel filtration and polyacrylamide gel electrophoresis. In both species two different T4 binding molecules were identified in whole serum and in Cohn fraction V preparations. Only one of these binding species demonstrated the characteristics of specific binding, i.e., high affinity for the hormone and binding site saturability. The mean (+/- SD) apparent association constants (ka) at 37 C for the specific binding proteins in whole rat and rabbit serum were 3.5 +/- 0.5 x 10(8)M-1 and 2.8 +/- 0.5 x 10(8)M-1, respectively. The mean T4 binding capacities of these proteins were 4.3 +/- 1.2 x 10(-6)M and 7.4 +/- 1.0 x 10(-6)M for rat and rabbit serum, respectively. Non-specific binding was due to serum albumin. Rat albumin (ka = 6.1 +/- 1.6 x 10(5)M-1, at 37 C) bound T4 significantly more strongly than did rabbit albumin (ka = 2.3 +/- 0.4 x 10(5)M-1, at 37 C) when it was assumed that there was only one T4 binding site/molecule of albumin. The ability of rat albumin to bind more T4 could also be explained by a greater number of T4 binding sites/molecule. Partial separation of the two T4 binding species was attained by gel filtration on Sephadex G200 columns. The specific binding protein of both rat and rabbit serum was eluted slightly later than the corresponding serum albumin. Polyacrylamide gel electrophoresis resulted in the separation of two distinct T4 binding peaks in the rat, but with rabbit serum clear separation of the two T4 binding molecules was not attained. In both species the specific binding protein migrated anodal to serum albumin.


Confirmation of pesticide residue identity: part IX. Organophosphorus pesticides. This paper describes a confirmation procedure for residue levels of dyfonate, dichlorofenthion and cyanox. After extraction and quantitation by flame photometric detector (FPD)-gas liquid chromatography (GLC), the organophosphorus pesticides are hydrolyzed in a 10% methanolic-potassium hydroxide solution. The phenolic hydrolysis products are then extracted from this solution and reacted with pentafluorobenzyl bromide to produce the pentafluorobenzyl (PFB) ether derivatives. These PFB ethers are subsequently cleaned-up and fractionated on a silica gel micro-column and analyzed by electron capture (EC)- GLC.


Inhibition of lobster-muscle arginine kinase by homologous antibodies and study of an antibody population directed against a tyrosine-containing antigenic structure. The effect of highly inhibitory rabbit antibodies on the enzymic activity of lobster-muscle arginine kinase has been studied.  100% inhibition was obtained at a 30-fold molar excess of antibody.  The guanidine substrate L-arginine did not protect the enzyme from subsequent inhibition by its antibodies.  On the other hand, the metal-nucleotide substrate Mg-ATP, or the substrate analog Mg-AMP, protected about 50% the enzyme activity, the extent of hte protection being irrespective of hte presence or of the absence of L-arginine and of the value of the molar ration Ab/Ag/  An antibody population directed against the antigenic determinant containing the essential tyrosine residue of the enzyme has been isolated by the affinity chromatography.  No inhibitory acitivity was found in that fraction.  Most of the inhibitory activity was found in the remaining antibody populations...


The electrophoretic mobility of serum lysozyme. The electrophoretic mobility of serum lysozyme in 2 patients with raised enzyme levels was identical to that of gamma-globulins. Similar mobility was observed after incubation of lysozyme and normal serum. Incubation with one hypogammaglobulinemic serum showed that lysozyme could also acquire alpha2 mobility.


Physiological effects of the meteorologically abnormal year 1974 observed in healthy males in the Netherlands and southern Norway. Convincing evidence of biological effects during long-term abnormal weather conditions is very rare because usually no continuous series of physiological data from the same animals or humans (either single or in groups) have been collected over many years during the period preceding the meteorological event. Since 1955, at the Biometeorological Research centre, Leiden, about 220,000 blood data were collected from healthy male blood donors at Leiden, The Netherlands, and about 110,000 data at Oslo, Norway, since 1966. It was found that during the meteorologically abnormal year of 1974, which was abnormal in many parts of the world, both in Leiden and Oslo abnormal biological patterns occurred in blood sedimentation rate, haemoglobin, diastolic blood pressure, gamma-globulin levels, Piccardi p-test percentages and a few other biological parameters. A more complete monograph will be published at a later date.


Levels of beta-trace protein and lysozyme in human amniotic fluids. The levels of beta-trace protein and lysozyme were estimated in amniotic fluids from normal fetuses and from fetuses with neuraltube defects. The values of these proteins in normal amniotic fluids were found to be similar to those detected in fetuses with anencephaly and spina bifida. The levels of lysozyme were shown to be correlated with gestational age.


Interrelations of matrix lipids, vesicles, and calcification. When calcifying tissues were extracted with hot pyridine or hot benzene and then decalcified and stained with Sudan black B, the areas where mineralization was being initiated stained strongly, the rest of the calcified tissues being unstained. Histochemical methods showed that lipids were responsible for the staining. They were isolated biochemically and found to be phospholipids, very resistant to extraction before decalcification of the tissues and consisting predominantly of phosphatidyl serine and phosphatidyl inositol. It was proposed that these phospholipids were active at nucleating sites in apatite crystal formation, since it is known that phosphatidyl serine binds calcium strongly. It has been shown that phosphatidyl serine is present in matrix vesicles.


Alcoholic hepatitis. Cell-mediated immunological response to alcoholic hyalin. Immunological reactivity in alcoholic hepatitis has bben attributed to alcoholic hyalin, the histological hallmark of this disease. A purified isolate of alcoholic hyalin with electron microscopic, biochemical, and serological characteristics documented previously was added to lymphocytes from healthy subjects and patients with alcoholic hepatitis or other hepatic disorders. Production of migration inhibition factor (MIF) in response to this material was used as an index to lymphocyte reactivity. MIF was significantly increased in lymphocytes obtained from patients with alcoholic hepatis, as compared to the healthy controls (P less than 0.001), and persons with other liver diseases (P less than 0.005). These observations indicate that immunological hyperreactivity to alcoholic hyalin occurs in patients with alcoholic hepatitis; such activity may be of key importance in the pathogenesis or sequelae (or both) of this disease.


[Influence of hypertensive substances on the kallikrein-kinin system of the blood]. It is shown that following a one-time intravenous administration to rats of ethyron, epinephrine and pituitrin (in doses causing hypertension) the amount of the blood plasma kininogen is more than halved with concurrently increasing caseinolysis. A two-day long introduction of these substances activates prekallicrein and brings down the level of the kallicrein inhibitors, whereas a 10 and 20-day coursewise administration of these compounds intensifies the synthesis of some components in the kallicrein-kinine system.


Hepatitis BS antigen, malaria titers, and primary liver cancer in South Vietnam. A total of 306 individuals from South Vietnam were studied: 61 had a diagnosis of primary liver cancer (38 had a tissue diagnosis, and 23 had a clinical diagnosis and a positive alpha-fetoprotein); 9 had viral hepatitis; 101 were hospitalized patients (60 with various other forms of liver disease and 41 without liver disease); 94 were blood donors; 29 were drug users, and 12 were medical students. Alpha-fetoprotein was present in 45 of 61 (74%) of those with a diagnois of primary liver cancer (PLC) and in none of the other patients. Using immunoelectroosmophoresis, hepatitis BS antigen (HBSAg) was found no more frequently in those with PLC than in the other groups studied. In contrast, using a radioimmunoassay technique HBSAg was present 3 to 8 times as frequently in the PLC patients as in other subjects without viral hepatitis. There was a close relationship between the presence of alpha-fetoprotein and HBSAg in the patients with PLC. Malaria seropositivity rates were no different in the PLC groups than the other groups. It appears that in South Vietnam PLC is associated with an increased frequency of HBSAg.


[Studies on juvenile chronic hepatitis]. Follow-up study of 40 children suffering from chronic hepatitis. The diagnosis was made by liver needle biopsy with the Menghini method, when clinical signs or laboratory data of liver disease had lasted for more than 6 months. 24 patients showed the histological pattern of the aggressiv type of chronic hepatitis according to the definition of the European Association for the Study of the Liver (1968). In this group only 5 children had autoantibodies in the serum (so-called lupoid hepatitis). The HBAg positive courses played the most important part in the chronic persistent group as well as in the aggressive one. According to literature only the patients with the aggressive type have been treated with prednison, because chronic persistent hepatitis has a good prognosis without any treatment. In nearly all cases high transaminases and gammaglobulin levels decreased during the treatment with prednison, whereas the histological signs of inflammation seldom changed. Cirrhosis of the liver has developed in 2 HBAg positive patients of the aggressive group, who had not consequently received their daily dose of prednison.


The role of trypsin in the pre-treatment of chromosomes for Giemsa banding. The role of trypsin in the elicitation of G-banding on human chromosomes was studied in two separate laboratories. Enzyme activity and ability of trypsin to chelate calcium were manipulated by dilution of the treatment solution, and by inhibition with diisopropylphosphofluoridate, diphenylcarbamyl chloride, or soybean trypsin inhibitor. In all cases, chromosomes were affected in proportion to the enzyme activity of the treatment solution rather than the ability of the solution to bind calcium. It is concluded that calcium chelation is not sufficient to explain G-banding trypsin, but that proteolytic activity is required.


Cytochemical study of pseudoisocyanine stained human chromosomes. Human meiotic and mitotic chromosomes were studied with N-N' diethyl pseudoisocyanine stain. Following methylation and oxydation, the staining allowed microscopic observation of slides with both monochromatic light and fluorescence. In addition, stained preparations can be permanently conserved. Preceeded by diverse methods of chromosome denaturation or 5-BUDR incorporation, PIC lends itself to a large number of banding techniques. Cytochemical study of stained chromosomes demonstrated a certain PIC affinity for DNA although tests performed do not exclude the possibility of PIC reaction with certain proteins.


Inhibition of anaphylactic histamine release by Forssman antiserum. I. Characteristics of the reaction and inhibitor. Forssman antiserum produced in rabbits immunized with sheep erythrocyte stromata was found to contain  an IgG antibody which inhibited both passive anaphylactic sensitization of guinea-pig lung and also histamine-releasing activity of soluble immune complexes. This Forssman antibody did not itself cause histamine release or depletion of lung histamine stores. The IgM haemolysin component of the Forssman antiserum was not associated with inhibitory activity. The inhibition by the IgG Forssman antibody differed from that of normal rabbit gammaglobulin both in its irreversible character and in being absorbed by sheep erythrocytes. The inhibitory antibody had no effect on the histamine-releasing activity of compound 48/80, anaphylatoxin or reversed anaphylaxis. It was concluded that IgG Forssman antibody probably blocks the tissue receptor(s) for anaphylactic antibody in guinea-pig lung.


Identification of sialic acid in polysaccharide antigens in group B Streptococcus. By means of paper chromatographic techniques with purified, sialic acid-containing, bacterial antigen standards, sialic acid, as well as galactose and glucosamine, have been identified as components of each type-specific antigen of group B Streptococcus. Rhamnose was not detected in the group B antigen isolated by the classical HCl and heat extraction method.


Correlation between fluorescent antibody detection of hepatitis B core antigen in liver biopsies and the presence of e antigen in serum. HBc Ag was detected by fluorescent antibody in liver biopsies taken from chronic hepatitis B surface antigen (HBs Ag) carriers having e antigen in their sera. HBc Ag could not be detected in liver biopsies from similar individuals having e antibody in their sera.


Immunochemical analysis of distinct cellular antigens isolated from a nontypable bovine strain of group B streptococci. Four immunologically distinct antigens were isolated from a bovine strain of group B streptococci, designated 14 Mi. Chemical analysis indicated that two of the cell surface antigens consisted of glucose, galactose, and glucosamine, whereas the other cell surface antigen, an acidic protein, contained a predominance of aspartic acid, glutamic acid, and alanine. A cell wall-associated triheteroglycan consisting of galactose, glucose, and glucosamine was isolated from strain 14 Mi by 10% trichloroacetic acid. Immunochemical studies suggested that this glycan is type specific and consists of an immunodominant alpha-linked galactosyl-glucose disaccharide.


Neutral proteases of human PMN leukocytes with kininogenase activity. A neutral protease with kininogenase activity was isolated from human polymorphonuclear (PMN) leukocytes by cation exchange chromatography and gel filtration. The protease appears heterogeneous by cation exchange chromatography, isoelectric focusing and cationic disc gel electrophoresis, but homogeneous by gel filtration, sucrose density gradient ultracentrifugation, SDS-disc gel electrophoresis and immunoelectrophoresis. By carrying out the electrophoresis of the protease in acrylamide gels of varying concentrations, it was shown that they represent charge isomers. The protease was stable at pH 4-10, but labile to heat, being almost completely inactivated when incubated for 30 min at 70 degrees C. It exhibited proteolytic activity between pH 5 and 9, being maximal at 7.5-8.5. The molecular weight of the PMN protease was estimated to be about 20,000 daltons by gel filtration in aqueous buffer and about 26,000-28,000 daltons by SDS-disc gel electrophoresis and gel filtration in Sepharose 6B in the presence of the dissociating agent guanidine HCl. Its sedimentation coefficient was about 2.7S. Corresponding to the charge heterogeneity, by isoelectric focusing, the kinin-generating and esterolytic activities of the PMN granule lysate focused between pH 6.0 and 11.5, whereas the isolated PMN protease focused between 10.0 and 11.8. With respect to kinin generation, caseinolysis, and alanine esterase activity, the protease was inhibited by DFP and certain chloromethyl ketone inhibitors, as well as the plasma protease inhibitory a1-antitrypsin, a2-macroglobulin and antithrombin III. Both bradykinin and a methionyl-lysyl-bradykinin-like peptide were generated from highly purified kininogens by a lysosomal lysate containing the PMN protease. However, this assay was done with a crude enzyme preparation which contains an aminopeptidase capable of converting lysyl-bradykinin or methionyl-lysyl-bradykinin to bradykinin. When injected intradermally, the protease induced hyperemia, hemorrhage, and moderate enhancement of vascular permeability, but the mixture of the protease and kininogen induced a marked enhancement of vascular permeability.


A receptor for antibody on B lymphocytes. III. Relationship to immunoglobulin and ia determinants. The physicochemical structure of the receptor for antibody (FcR) on B cells and its interrelationship with Ig and H-2 gene complex associated antigens were examined. FcR were found to be sensitive to treatment with phospholipase C and pronase, but resistant to neuraminidase, phospholipase A and chymotrypsin. They would therefore appear to be composed of phospholipoproteins. Several lines of evidence indicated that FcR and Ig receptors were discrete entities: thus, FcR (1) were resistant to chymotrypsin; (2) capped independently of Ig, as demonstrated by means of Fab fragments of anti-Ig, and (3) were closely associated with at least some Ia determinants, which are known to be distinct from Ig determinants. The relationship between FcR and H-2 gene complex associated antigens was confirmed by demonstrating inhibition of binding of aggregates by anti-Ia serum and vice versa. If, however, FcR were capped, anti-Ia serum applied under non-capping conditions was still found to bind diffusely to the great majority of B cells. Although this could be explained in part by the presence of residual FcR, some Ia determinants appeared to be distinct from FcR. The finding of residual FcR after capping with aggregates or immune complexes implied that FcR are a more integral part of the cell membrane than Ig receptors and could therefore act as proreceptors for the latter. Consistent with this was the demonstration of a significant polar distribution of Ig on B cells capped for FcR and then labelled under non-capping conditions with anti-Ig.


Serological investigations in 15 cases of bird fanciers disease. Serological studies in 15 patients with bird fanciers disease are reported. Quantitative analysis of serum proteins showed elevation of IgG in almost every case. High IgE was observed in three patients with isolated asthmatic reactions. In four patients alpha1-antitrypsin was transiently diminished. Two of these had heterozygous deficiency of this protein (genotype MS and MZ). Precipitins against avian sera and avian droppings were found in all patients, but also in some exposed and nonexposed controls. Cross-reactions to avian antigens from different species were observed quite regularly in patients but not in controls. One case is presented suggesting that these cross-reactions may be of clinical importance. Precipitins against other inhaled antigens such as Aspergillus fumigatus, cereals or Micropolyspora faeni were observed significantly more frequently in patients than in controls. These precipitins showed no cross-reactions with those against avian antigens.


Inhibition of 3H-thymidine incorporation into human lymphocytes by dialyzable material of streptokinase- streptodornase. Using the 3H-thymidine incorporation test, the lymphocyte-stimulating activity of streptokinase-streptodornase (SK-SD, Varidase) and its fractions were evaluated. The dose-response curves of all the preparations except the third fraction had two peaks with the maximum around 5 and 40 mug. The dialyzable fraction of Sk-SD (D-SK-SD) occupying approximately half of the original sample had almost no lymphocyte-stimulating activity; and conversely, it had a potent inhibitory effect on the 3H-thymidine incorporation into lymphocytes that normally occurs with non-dialyzable SK-SD (ND-SK-SD), PPD and PHA-P. Therefore, it will be desirable to remove the dialyzable fraction from the commercial SK-SD preparation when it is used for the determination of the capacity of cellular immunity against tsk-sd.


Measurement of dengue virus-specific double-stranded ribonucleic acid in infected primate cell extracts by microquantitative complement-fixation methods. Microquantitative complement-fixation methods have been applied to the detection and quantitation of double-stranded RNA (dsRNA) in cytoplasmic extracts of uninfected and dengue virus-infected primate cell lines. LLC-MK2 and PGLC-33H cells exhibited increased levels of dsRNA following dengue virus infection. Cytoplasmic extracts of infected and uninfected LLC-MK2 cells subjected to density gradient analysis showed that increased levels of dsRNA were associated with structures believed to be virus-specific. Primate cell dsRNA and dengue dsRNA preferentially reacted with rabbit anti-rIn-rCn, while reovirus dsRNA and rAn-containing synthetic duplexes preferentially reacted with anti-rAn-rUn.


Inhibition of allergic reactions by chromoglycate and by a new anti-allergic drug (10-chloro-1,4,6,9-tetrahydro-4,6-dioxopyrido (3,2-g) quinoline-2,8-dicardoxylic acid). I. Activity in rats. A new inhibitor of IgE-mediated allergic reactions has been studied in rats. This new inhibitor is chemically quite different from cromoglycate and shows qualitative and quantitative advantages over it. This new inhibitor possesses no bronchodilator or end-organ antagonism activity, but as cromoglycate acts by inhibition of mediator release. Multiple doses in rats lead to tachyphylaxis. This effect is reversed both by decreasing the first dose and increasing the time between doses.


Studies in experimental eosinophilia. XI. Dependence of eosinophilia, apparently induced by histamine, on acidity. A solution of histamine in saline elicited eosinophil response in both lymph nodes and the blood stream. The lymph node response was not inhibited by the antihistamine, mepyramine maleate, but was inhibited by puromycin. Both the lymph node and the systemic response were essentially abolished when the pH of the solution (5.0 and 4.7, respectively) was brought to neutrality by phosphate buffer. Serum could bring the pH of histamine to neutrality or higher, but the histamine-serum mixture retained in vivo activity, unless the histamine was buffered before addition to the serum. When normal guinea pig gamma-globulin was incubated with histamine, a nondialyzable factor was generated which had the same activity as histamine. Thirteen different amino acids induced lymph node eosinophilia and each was acid in saline solution; when one of them, trytophane, was brought to neutral pH, its activity was considerably reduced. During the period when the circulating level of eosinophils was rising, there was no detectable fall in the bone marrow stores of mature of immature eosinophils. Thus, the activity of histamine, vis-a-vis eosinophils, is nonspecific and is attributable to its acidity. The data suggest that histamine, in the acidic state, generates autoantigens, which, in turn, lead to antibody-mediated eosinophilia in lymph nodes.


EBV-determined nuclear antigen (EBNA)-positive cells in the peripheral blood of infectious mononucleosis patients. After removal of SRBC rosette-forming T-cells from the peripheral blood, the residual, largely B-lymphocyte fraction of five infectious mononucleosis patients was found to contain 0.5-2% blast cells, positive for the EBV-determined nuclear antigen (EBNA). There was a rough parallelism between the presence of large lymphoblasts in the hematological smear, EBNS-positive large blasts in the B-cell fraction and the ability of the T-cell fraction to exert an EBV-specific lymphocytotoxicity on established cell lines in vitro. EBNA-positive B-cells and EBV-specific killer T-cells disappeared after the acute phase of the disease.


Tumor-bound immunoglobulins. Evidence for the in vivo coating of tumor cells by potentially cytotoxic anti-tumour antibodies. The experiments described herein were designed to determine whether part of the Ig coat of tumor cells consists of specific anti-tumor antibodies. It was demonstrated that the inoculation of polyoma virus-induced sarcoma cells (SEYF-a) into syngeneic A.BY mice stimulates the production of cytotoxic antibodies against the tumor-cell population. The level of these antibodies, which was undetectable during the first week after transplantation, increased markedly during the second week, and remained high thereafter. Following the increase in cytotoxic antibodies in the serum, a cell-bound potentially cytotoxic antibody was detected on the tumor cells by testing their sensitivity to rabbit complement. The increase in cell-bound, potentially cytotoxic antibody followed the kinetics of the increase in serum antibody during the second week after transplantation and was inversely correlated to the amount of free antigens on the cell surface. These antigens, responsible for the sensitivity of the cells to a syngeneic hyperimmune cytotoxic antiserum, became non-available for the cytotoxic antibodies during propagation of the tumor cells. Cells from a tumor propagated for 3 weeks could not compete for anti-tumor antibodies with cells propagated for 1 week. Yet it was possible to increase the antigenic capacity of cells from an old tumor by a treatment that would cause the release of tumor-associated Ig. Cytotoxic anti-SEYF-a antibodies could be dissociated from tumor cells propagated in vivo by methods causing dissociation of antigen-antibody complexes, and detected in tumor eluates.


Persistence of Epstein-Barr viral nuclear antigen (EBNA) in cells entering the EB viral cycle. It was shown by double immunofluorescence studies that Epstein-Barr viral nuclear antigen (EBNA) was preserved in EBV-infected cells after they had entered the productive viral cycle, as signalled by the appearance of the early antigen (EA)complex. A nuclear component of the EA comples could be clearly distinguished from EBNA with regard to antigenic specificity.


[Histochemical demonstration of pancreatic B-cells by means of the colloidal iron (Hale) reaction (author's transl)]. The stainability of B-cells in islets of Langerhans by means of colloidal iron reaction has been examined using two standard modifications (Graumann resp. Mowry) of the original Hale-reaction. After previous oxidation of the sections with performic acid a strong and selective staining of B-cells was obtained by the use of a colloidal iron reaction based upon Graumann's method. With Mowry's technique B-cells remained unstained. The demonstration of B-cells using a performic acid-colloidal iron reaction has been compared with methods of known selectivity (Aldehyde Fuchsin, Dichlorpseudoisocyanine). By restaining procedures it could be shown that with the three methods the same type of cell i.e. B-cell is stained.


The histochemical application of dansylhydrazine as a fluorescent labeling reagent for sialic acid in cellular glycoconjugates. A new method for the fluorescent staining of stalic acid-containing glycoconjugates in fixed tissues is described. The procedure uses mild periodate oxidation, followed by condensation with dansylhydrazine and reduction of the hydrazones to hydrazines. The specificity of the reaction for sialic acid is tested on model glycoconjugates. The procedure gives superior resolution in comparison to the standard periodate Schiff procedure for cellular carbohydrates.


Glycoprotein metabolism in the hypothalamus of rat: significance of glial cells. Incorporation of 3H-fucose injected into the CSF was studied in hypothalamus and hypophysis of rats by biochemical and autoradiographic techniques. The concentration of radioactivity was determined as a function of time from 10 minutes to 28 days in the acid soluble and the acid precipitable fraction of homogenates. The incorporated activity was localized by light microscopic autoradiography in several regions of the hypothalamus. The labelled proteins were transported to the pars nervosa at a velocity of 1.1 to 3.3 mm/hr, i.e. in the fast component of the axoplasmic flow. Differences were found between hypothalamic and pars nervosa tissue regarding rates of increase and maximum concentrations of protein-bound radioactivity. Values for both parameters were about 6 times higher in hypothalamus than in pars nervosa. On the basis of the autoradiographic observations it is concluded that this difference may be due to a considerably higher synthesis rate of glycoproteins in glial cells than in neurons.


A comparison of four quantitative cytochemical methods directed toward demonstration of DNA. Populations of nuclei isolated from mouse brain tissue were stained by the following cytochemical methods considered stoichiometric for DNA: (1) the Feulgen reaction; (2) gallocyanin-chromalum after RNase; (3) pH 4.0 methylene blue after RNase; and (4) methyl green used in the presence of 2M magnesium chloride. Replicate preparations to be stained with gallocyanin-chromalum, methylene blue, and methyl green were acetylated prior to staining. All of these groups were examined by high-resolution scanning microspectrophotometry. The results indicated that of the methods examined, the Feulgen reaction, gallocyanin-chromalum used without prior acetylation, and methylene blue used with prior acetylation were the most useful in revealing differences attributable to variability in chromatin organization. The greatest variability in total extinction measurements was observed in acetylated, methylene blue-stained nuclei, while the least variability was observed in nuclei stained with methyl green in the presence of 2 M magnesium chloride. Acetylation produced different effects on dye-binding in different groups. It greatly increased binding in nuclei stained with methylene blue; it reduced binding in the methyl green-2 M magnesium chloride series.


Electron microscopic localization of monoamine oxidase in the rat liver. Two methods have been employed to localize monoamine oxidase activity in the cells of rat liver, using either 2-(2'-benzothiazolyl)-5-stryl-3-(4'-phtalhydrazidyl) tetrazolium chloride (BSPT) or ferricyanide as electron acceptor. With both methods monoamine oxidase activity was found both in the inner and the outer mitochondral membrane, although the outer membrane appeared the most probable location. In addition the BSPT method but not the ferricyanide method, revealed monoamine oxidase activity in the endoplasmatic reticulum. The results obtained by the two methods have been compared and are discussed in view of available biochemical data on monoamine oxidase.


Glycoproteins in membranes of secretory granules of the anterior pituitary gland. Rat anterior pituitary glands were examined by electron microscopy after staining with five different histochemical stains. Histochemical reactions were observed in the cell coat, cell membrane and the membrane surrounding the secretory granules in all anterior pituitary cells following staining with phosphotungstic acid (PTA), chromic acid and PTA, the periodic acid-thiosemicarbazide-silver protein method (PA-TSC-SP) of Thiéry, ruthenium red and concanavalin A. The staining was abolished when the sections were preincubated with pronase, neuraminidase or trypsin and subsequently exposed to PTA, chromic acid and PTA or PA-TSC-SP. The possible functional role of the glycoproteins present in the membrane surrounding the secretory granules is considered.


Further observations on the chemistry of pararosaniline-Feulgen staining. Pararosaniline-Feulgen staining of cells in suspension produces nucleus- and chromatin-specific fluorescence as well as color. Experiments were designed to test postulated reaction mechanisms responsible for the fluorescent staining with the nonfluorescent pararosaniline. The reduction in fluorescent-staining intensity by pretreatment of cells with 2.2 x 10-2M K2S2O5 tends to rule out the alkysulfonic acid pathway; conditions favoring the formation of this intermediate reduce staining intensity. The fluorescence enhancement, observed when cells stained in pararosaniline without K2S2O5 are post-treated with K2S2O5, suggests that there is an initial Schiff-base linkage between pararosaniline and an aldehyde of hydrolyzed DNA, and that this linkage is stabilized in the presence of K2S2O5. Microspectrofluorometer measurements of cells stained at various pararosaniline concentrations in 2.2x10-2M K2S2O5, show that the fluorescence emission maximum ranges from about 627 nm at 3.1x10-3 M pararosaniline to about 604 nm at 3.1x10-5M. All of the employed staining protocols appear to produce the same fluorescent product, perhaps a heterocyclic pyronin analog formed from pararosaniline. Flow microfluorometric analysis of cells stained in suspension verified that the relative fluorescence intensity represents relative DNA content. Staining at reduced pararosaniline concentration (3.1x10-4M) reduces the coefficient of variation of the flow microfluorometric histograms, showing that maximum quantitation does not necessarily correlate with maximum staining intensity.


The use of a transesterification technique to distinguish between certain neuraminidase resistant epithelial mucins. The effects of potassium hydroxide and soidum methoxide treatments upon the Alcian blue staining and neuraminidase lability of certain neuraminidase resistant epithelial mucins have been studied. The results were interpreted as indicating that while the mucins of rat colon and rabbit Brunner's gland contain only 4-0-acetyl sialic acid, human colonic epithelial mucins may contain some sialic acid with esters at the C1 carboxyl group.


Production of specific antibodies to contractile proteins, and their use in immunofluorescence microscopy. I. Antibodies to smooth and striated chicken muscle myosins. Antibodies prepared against actomyosins can be shown to behave similarly, if not identically to more recently prepared antibodies against highly purified myosins. Details of the purification of the antigens, and of the production of antibodies to chick myosins from smooth gizzard muscle and from striated pectoral muscle are given. The antibody specificity appears to be directed against the heavy chains of the myosin molecules, since these antibodies specifically inhibit the myosin ATPase reaction, and since in situ staining of myosin polypeptide chains on an SDS gel using the antibodies in indirect fluorescence shows staining only in the heavy band region. Use of the antibodies in immunofluorescence microscopy suggest that the antibodies are tissue, but not species, specific. Example of their use in staining tissue sections are shown.


Light microscopic demonstration of myoid material in nuclei. During the development of configurational staining methods for proteins of the myosin-fibrin group, nuclei showed staining properties similar to those of myofibrils. This dye binding could be attributed to nuclear alpha-helical proteins. More recent chemical and electron microscopic studies demonstrated actomyosins in nuclei of various species. Possible roles of nuclear actomyosin in chromosome movements and condensation and in cell proliferation have been suggested. It seems therefore permissible to assume that the tannic acid-phosphomolybdic acid (TP)-Levanol Fast Cyanine 5RN method and similar technics visualize myosin in nuclei. Comparative studies of actomyosins from various sites indicated significant chemical an histochemical differences. It is therefore suggested that, in analogy to the different classes of collagens, there may be several subgroups of myosin which differ in their physico-chemical properties and sensitivity to fixation procedures and pathological conditions.


The histochemical demonstration of calcium with 8-hydroxyquinoline. A method for the histochemical demonstration of calcium using 8-hydroxyquinoline was investigated. It was found to give intense, highly selective staining of various pathologic and natural calcifications, and compared favourably with conventional procedures for calcium. Calcium oxalate and calcium pyrophosphate were not stained.


Sudden interruption of leaflet opening by ventricular contractions: a mechanism of mitral regurgitation. The motion of both mitral cusps and the presence of valvular regurgitation during ventricular contractions were investigated in seven experiments on dogs in which radiopaque markers had been sutured to the cusps and the valve annulus 1-32 wk before the studies. Cineangiograms of the left ventricle were obtained during ventricular ectopic beats, interposed throughout the cardiac cycle (20-99% of cycle length) and during induced variations in the P-R interval (0-200 ms). Mitral regurgitation was observed only during a) weak, early ectopic beats (peak pressure below 34 mmHg) which were incapable of closing the cusps and b) when ventricular contractions suddenly interrupted normal leaflet motion toward the ventricle, during three well-defined periods of diastole (diastolic valve opening, diastolic rebound, and atrial opening). Valve closure following sudden reversal of cusp opening was slow and the leaflets often did not arrive simultaneously at their closed positions. These findings suggest that sudden interruption of leaflet opening by ventricular contractions is an important mechanism of transient mitral regurgitation in the normal heart.


Inhibition of Ca2+ uptake into fragmented sarcoplasmic reticulum by antibodies against purified Ca2+, Mg2+-dependent ATPase. Rabbit antiserum was prepared against a partially purified Ca2+, Mg2+-dependent ATPase [EC 3.6.1.3] of the SR isolated from chicken skeletal muscle. The gamma-globulin fraction of antiserum contained antibodies which combined with the purified ATPase and the SR vesicles. Binding of the antibodies strongly inhibited active transport of Ca2+ ions into the SR, but not passive leakage of Ca2+ ions from the SR. The antibodies scarcely affected the ATPase activity.


Aglycosylantibody. Effects of exoglycosidase treatments on autochthonous antibody survival time in the circulation. Rabbit anti-hapten antibodies were purified by affinity chromatography and characterized immunochemical for in vivo studies of their blood clearance rate and organ distribution after treatment with various glycosidases. Following sequential removal of sialic acid, galactose, and N-acetylglucosamine with the appropriate cellulose-immobilized exoglycosidases, the antibody populations were recharacterized, radiolabeled, and introduced intravenously into the original animals. Using double radioiodine lables it was possible to demonstrate alterations in purified antibody survival times in the circulation and altered organ distribution after glycolytic cleavage. Removal of terminal sialic acid resulted in rapid blood clearance and enhanced localization of asialoantibody in the liver. Subsequent removal of penultimate galactose residues returned both antibody survival time in the circulation and organ distribution to near normal. Removal of subpenultimate N-acetylglucosamine moieties resulted in aglycosylantibody survival values which were intermediate between asialo- and asialoagalactoantibodies. Removal of the three saccharide also increased kidney localization. The results are evaluated based on current concepts of the biological roles of protein-linked carbohydrate and plasma glycoprotein survival time in the circulation.


On the fidelity of DNA replication. Enzyme activities associated with DNA polymerases from RNA tumor viruses. DNA polymerase from RNA tumor viruses ("reverse transcriptase") has been analyzed for activities which have been associated with other DNA polymerases. Homogeneous DNA polymerase from avian myeoblastosis virus catalyzes pyrophosphate exchange and pyrophosphorolysis. Pyrophosphate exchange is dependent on a template and is base-specific. With avian myeloblastosis virus DNA polymerase, ribonucleotide templates are more efficient for synthesis while deoxyribonucleotide templates are more effective for pyrophosphate exchange. Synthesis, pyrophosphate exchange, and pyrophosphorolysis were inhibited by the chelating agent 1,10-phenanthroline, suggesting that enzyme-bound zinc is required for each of these reactions. The pyrophosphate exchange reaction was also demonstrated with the DNA polymerase from a mutant of Rous sarcoma virus that possesses a temperature-sensitive DNA polymerase. The pyrophosphate exchange reaction with the mutant polymerase is temperature-sensitive which demonstrates that pyrophosphate exchange is indeed catalyzed by the viral DNA polymerase and that the same mutation effects both DNA polymerase and pyrophosphatase activity. Unlike Escherichia coli DNA polymerase I, the DNA polymerase from avian myeloblastosis virus fails to degrade polydeoxyribonucleotides or to convert deoxynucleoside triphosphates into monophosphates. This lack of hydrolytic activities in avian myeoblastosis DNA polymerase should facilitate kinetic studies on the mechanism of DNA synthesis by this enzyme.


On the fidelity of DNA replication. Lack of exodeoxyribonuclease activity and error-correcting function in avian myeloblastosis virus DNA polymerase. Homogeneous DNA polymerase ("reverse transcriptase") from avian myeoblastosis virus was assayed for exodeoxyribonuclease activity. The substrates were defined template-initiator complexes in which different radioactive nucleotides were present at the 3'-OH termini of the initiator. Even when the number of molecules of enzyme was equal to the number of initiator termini there was no significant release of radioactivity with any of the template-initiator combinations tested. Under similar conditions, the nuclease activity associated with either Escherichia coli or T4DNA polymerases rendered more than 90% of the initiator termini acid-soluble. The ratio of exodeoxyribonuclease activity to protein with avian myeoblastosis DNA polymerase is less than 0.003% of that obtained with E. coli DNA polymerase I. Furthermore, avian myeloblastosis virus DNA polymerase failed to excise mispaired terminal nucleotides in both the presence and absence of polymerization.


Resin hemoperfusion: a method of removing circulating thyroid hormones. The ability of an extracorporeal hemoperfusion system employing neutral Amberlite resin to bind thyroid hormone and to decrease circulating levels of triiodothyronine (T3), thyroxine (T4), and free thyroxine (FT4) was evaluated in dogs made thyrotoxic by the intramuscular administration of thyroid hormone. Since the resin column and tubing were charged with saline, the effects of hemodilution from this source on serum T3 and T4 was assessed by control perfusion through a column which did not contain any resin. After correction for hemodilution, the mean serum T3, T4 and FT4 decreased during 2 hours of resin hemoperfusion by 39%, 35%, and 46%, respectively. Hormonal clearance rates were calculated in two experiments and the estimated net hormone removed averaged 60.4 mug of T3 and 1990 mug of T4. Hematologic indices and routine chemistries did not change significantly in these dogs during the procedure except for a decrease in mean serum albumin concentration and an increase in mean serum glucose concentration. Hemoperfusion through this resin system seems to be a safe, effective means of decreasing serum T3, T4, and FT4 in thyrotoxic dogs and warrants evaluation for the treatment of thyroid storm in man.


Reagin-mediated asthma in rhesus monkeys and relation to bronchial cell histamine release and airway reactivity to carbocholine. Rhesus monkeys with persistent immediate-type cutaneous and respiratory responses (RR) to ascaris antigen (AA) were compared with rhesus monkeys with skin reactivity and no respiratory responses, and animals with no skin reactivity and no respiratory responses to inhaled antigen (NR). The RR group could not be distinguished from the nonresponding  (NR) group by the cutaneous skin test titers, serum, or respiratory secretion IgE concentration. Leukocyte histamine (H) release due to anti-IgE was similar with peripheral blood leukocytes and bronchial lumen mast cells (MC) from RR and NR animals. The RR group of animals could be distinguished from the NR group by their degree of sensitivity to inhaled carbocholine and H release from respiratory MC exposed to AA. The RR group demonstrates consistent, persistent respiratory responses suitable for immunologic, pharmacologic, and physiologic studies. Finally, it was found that the IgE concentration in respiratory secretions of rhesus monkeys was comparatively higher than in serum, evidence for IgE as a secretory Ig in the respiratory tract of this species.


Fractionation of cells on a discontinuous Ficoll gradient. Study of subpopulations of human T cells using anti-T-cell antibodies from patients with systemic lupus erythematosus. Patients with active systemic lupus erythematosus (SLE) had a decrease in a subpopulation of cells (fraction D) when peripheral blood lymphocytes were separated on a discontinuous Ficoll gradient. Preincubation of SLE cells at 37 degrees C for 30 min led to a marked decrease in this fraction, composed primarily of thymus-derived (T) cells. Supernates of such preincubations were found to cause a reduction in fraction D cells from normal humans. The active factor in the supernate was found to be an IgG antibody. Similarly, serum from patients with active SLE produced a reduction in fraction D cells from normal donors. This activity was also found in the IgG fraction, and could be absorbed with a pure T-cell population. Depletion of macrophages and complement did not reduce the SLE anti-T-cell antibody-mediated loss of cells from fraction D; however, heat-aggregated human gamma globulin led to impairment of the reaction. These findings suggest that antibody-dependent direct lymphocyte-mediated cytotoxicity may play a role in T-cell lymphopenia of SLE. It was further noted that the SLE anti-T-cell antibodies, in contrast to rabbit antihuman thymocyte serum, recognized fraction D cells but not fraction E cells from normals. Since both fractions are largely T cells, it appeared that the SLE serum was directed against cell-membrane antigenic determinants present on fraction D T cells, which were absent or reduced in quantity on fraction E T cells. Thus, evidence was presented indicating the presence of at least two subpopulations of cells in man. This was supported by differential absorption of the anti-T-cell sera with fractions D and E.


Experience with the sodium sulphate-Alcian Blue stain for amyloid in cardiac pathology. The sodium sulphate-Alcian Blue (SAB) method, which stains amyloid green, was evaluated in 220 hearts from elderly patients. The technique proved superior to the Congo red, crystal violet, and thioflavine T methods used either singly or as a battery for the demonstration of cardiac amyloid. Amyloid was easily identified under the X3 objective, even in small amounts. A few non-amyloid components stained varying shades of green but were easily distinguished on morphological grounds. No false positive or equivocal reactions occurred, and in particular elastic laminae and paravascular connective tissue were not tinctorially confused with amyloid. The SAB stain is technically simple and consistently reproducible, and no special light source is required for examination. An additional advantage in cardiac pathology is the simultaneous demonstration of any fibrosis, basophilic myofibre degeneration, tissue mast cells and mucoid degeneration of valves present.


A method for the differential determination of plasma antithrombins. A method for the differential determination of plasma antithrombins, antithrombin III and alpha2 macroglobulin, is described. The method is based on the selective inactivation of plasma alpha2 macroglobulin by treatment with 0-1 M methylamine for 10 minutes at 37 degrees C and on the observation that antithrombin III and alpha2 macroglobulin inhibited in defibrinated plasma low concentrations of thrombin without mutual interference and according to pseudo-first order reaction. In healthy subjects antithrombin III was shown to account for about 70% of the total antithrombin activity. But in patients with liver cirrhosis, where low levels of total antithrombin activity were observed, the relative contribution of antithrombin III was found to be noticeably lower.


Effect of student academic ability on learning from programed text, slide-tape, and lecture. The results of this study demonstrate that the programed text, slide-tape, and lecture do not differ greatly in effectiveness at any ability level. This fact eliminates the potential nightmare of having to assign dental students to different teaching formats according to academic ability. Future research should concentrate on the long-range effect of self-instructional materials, especially for the development of the effective study habits and self-reliance, which may be particularly important for low academic ability students. It is possible that academic ability is not the most important factor in matching students to teaching formats. Perhaps student personality characteristics will yield more useful guideposts for tailoring teaching to the student's best advantage.