Effect of bleomycin on [3H]Thymidine 5'-Triphosphate incorporation into host liver and hepatoma nuclei. The effect of bleomycin on [3H]thymidine 5'-triphosphate ([3H]TTP) incorporation into isolated sucrose nuclei from host liver and Morris hepatomas has been compared. Bleomycin stimulates [3H]TTP incorporation 13-fold in host liver and hepatoma 16 nuclei, 8-fold in hepatoma 7800 nuclei, and 3-fold in hepatoma 7777 nuclei. Differences in the nuclear membranes are not responsible for the different response of the nuclei. Nuclei, denuded of their membranes by Triton X-100 treatment, give similar results to sucrose nuclei. Analysis of DNA extracted from liver or hepatoma nuclei incubated with bleomycin indicates that bleomycin produces scissions in the nuclear DNA and that some repair synthesis takes place. Incubation of nuclei with 111indium-labeled bleomycin shows an equal binding capacity of liver and hepatoma nuclei for bleomycin. Bleomycin also stimulates incorporation of [3H]TTP in a system using chromatin or calf thymus DNA as primer. Host liver or hepatoma chromatin incubated with a DNA polymerase extracted from normal rat liver nuclei is stimulated approximately to the same extent by bleomycin. When DNA polymerase extracts from host liver and hepatoma nuclei are assayed with calf thymus DNA as primer, bleomycin has a greater stimulatory effect on [3H]TTP incorporation with host liver DNA polymerase than with hepatoma DNA polymerase in the system. We suggest that a defect in the repair system in hepatoma nuclei is responsible for the relatively lower response to bleomycin.


Specific enhancement of tumor growth and depression of cell-mediated immunity following sensitization to soluble tumor antigens. W/Fu rats inoculated s.c. with less than or equal to 5 x 10(7) syngeneic (C58NT)D (Gross virus-positive) lymphoma tumor cells normally develop a palpable tumor which reaches its maximum size (12 to 14 mm) at 6 to 8 days and is subsequently rejected by 10 to 12 days. However, rats previously sensitized with soluble tumor antigens from (C58NT)D cells prior to (C58NT)D tumor inoculation demonstrate a significant enhancement of tumor growth (the tumor reaches up to 26 mm and is rejected by 16 to 18 days). This enhancement persisted in antigen-treated rats that continued to receive soluble antigen after tumor inoculation. The in vivo enhancement coincided with a significant in vitro depression of cell-mediated cytotoxicity [assessed with 51Cr-labeled (C58NT)D target cells and peripheral blood leukocytes]. The observed tumor enhancement was specific, inasmuch as presensitization to either soluble tumor antigens from WR6 (Gross virus-negative) tumor, syngeneic to W/Fu rats, or to soluble antigen from W/Fu spleen cells had no enhancing effect on (C58NT)D tumor growth. Interestingly, sensitization to soluble tumor antigen alone did not elicit detectable cell-mediated immunity, cytotoxic antibody, or serum-blocking activity to the (C58NT)D tumor. We conclude that sensitization to soluble tumor antigens specifically impairs the immune apparatus normally acting in tumor rejection. This impairment appears to act primarily at the induction phase of the immune response.


Radioimmunoassay of bleomycin.

Radioimmunoassay of bleomycin. A radioimmunoassay for bleomycin has been produced using 125l-labeled bleomycin and antisera raised in rabbits against a carbodiimide-catalyzed bleomycin-bovine serum albumin conjugate. 125l-Labeled bleomycin was synthesized by direct iodination of the drug using the chloramine-T technique. The standard curve of the assay was linear on a logit-log plot and the lower limit of sensitivity was 250 pg bleomycin sulfate. A mean recovery of 102.6% (+/- 3.3% S.E.) was obtained using bleomycin added to normal sera. No significant decrease in bleomycin immunoreactivity was observed following 24 hr incubation of the drug in serum at 37 degrees. The radioimmunoassay was also suitable for measuring bleomycin in the presence of other drugs since the assay was not significantly affected by the other antineoplastic agents tested. The sensitivity and specificity of the radioimmunoassay for bleomycin should provide a new means for pharmacokinetic and toxicity studies of bleomycin.


The disposition and pharmacokinetics in humans of 5-azacytidine administered intravenously as a bolus or by continuous infusion. The disposition of 5-[4-14C]azacytidine, administered i.v. as a bolus or continuous infusion, was studied in cancer patients. After bolus, plasma 14C levels exhibited as multiphasic disappearance pattern; half-life (t1/2, beta phase) = 3.4 to 6.2 hr. Of 14C in plasma, less than 2% was associated with 5-[4-14C]azacytidine 30 min after dose. The ratios of 14C levels were: red cells/plasma, approximately 0.8; leukocytes/plasma, 1.1 to 2.3; nucleic acids/leukocytes, 0.2 to 0.43; sputum/plasma, 0.05 to 0.17. Urinary excretion (3 days) accounted for 73 to 98% of 14C, LEss than 1% in feces. The relative concentration of 5-azacytidine in plasma with continuous infusion stayed higher than with bolus; urinary excretion was similar. Fewer side effects were observed with continuous infusion than with bolus. The stability of 5-azacytidine was determined in various media at several temperatures by thin layer chromatography and nuclear magnetic resonance. At 20 degrees in Ringer's lactate (pH 6.2), the t1/2 was 94 to 100 hr. Stability increased with lowering of temperature and pH. From our data we conclude that 5-azacytidine should be given by continuous infusion rather than as a bolus.


Some effects of bleomycin on the proliferation, maturation time and protein synthesis of hairless mouse epidermis. Hairless mice were given 2 mg Bleomycin i.p. in 1-0 ml saline on two successive days. By a stathmokinetic method, by micro-flow fluorometry and by autoradiography certain kinetic parameters were measured during 10 days after the last injection. Cell counts were made and the turnover time of the differentiating cells estimated. Protein synthesis was estimated by the uptake of radioactive histidine, and dry cell mass measured by weighing. Bleomycin affected cell proliferation in the epidermis by depressing biphasically both the number of cells in, and the passage of cells through, the cell cycle phases: S, G2 and M, most probably by directly affecting late G1 cells and cells in mitosis. The time between the two minima of depressed DNA synthesis corresponded to the mean generation time of the basal cells. Histidine uptake and dry cell mass were slightly affected, but the turnover time of the differentiating cells was prolonged. Bleomycin thus had a strong long-lasting inhibitory effect on epidermal cell proliferation and a marked inhibitory effect on epidermal cell maturation in mice.


Stimulated DNA synthesis in livers and kidneys induced to proliferate associated with unchanged thymidine and thymidylate kinase activities. Enhanced mitotic activity and stimulated DNA synthesis associated with unchanged activity of thymidine and thymidylate kinases were observed in mouse kidneys induced to proliferate by intracardiac injection of lead acetate, and in rat livers following repeated administration of 5-azacytidine. On the other hand, the enhanced thymidine kinase activity evoked by L-tryptophan given by intubation at later stages of liver regeneration was paralleled by the enhanced incorporation of thymidine into DNA only to a very small degree.


Further investigations on the affinity chromatography of unlabeled and labeled cortisol-binding immunoglobulin. A specific method for the isolation of cortisol-binding immunoglobulin from rabbit antiserum by affinity chromatography on sepharose-coupled haptene (sepharose-cortisol) is described. This isolation technique has the advantage that no nonspecific gamma-globulin is present in the eluate of the column; previous absorption of anti-BSA-globulin can be omitted. The nature of the isolated immunoglobulin is determined by immuno-electrophoresis. The same isolation procedure can be used for the purification of radioactive antibody for radioimmunometric assay after labeling the immunoglobulin with iodine-125.


[Subcutaneous mycotic cysts in Madagascar (Histological aspects. Diagnostic value of the Gomori-Grocott stain. Associations with pheo-sporotrichosis)]. The authors report three cases of mycosic subcutaneous cysts and study the problem of the phaeo-sporotrichosis, a cystic lesion due to dematies brown fungi which are often introduced under the skin by a sting. Because of the absence of brown pigmentation of the fungi present in the three cases reported, the authors propose to group these various cysts under the name of "syndrome of the mycosic cysts" a part of which only being constituted by the phaeosporo trichosis. They emphasize the importance to draw surgeons' attention on such lesions, a priori clinically banal, and pathologists' attention on Gomori-Grocott staining in tropical countries.


beta 2-microglogulin levels in cancerous and other disease states. Serum beta2-microglobulin levels were measured, by radioimmunoassay, in patients suffering from a variety of benign and malignant clinical disorders. Elevated beta 2-microglobulin values were found in neoplastic and non-neoplastic disorders affecting a variety of organs. The most striking increases in beta 2-microglobulin are found in the plasma cell dyscrazias and several solid tumors, particularly those affecting the lung. Lymphoid neoplasms demonstrate a spectrum of changes of serum beta 2-microglobulin. At the one end of this spectrum were the plasma cell tumors, which show a high incidence of raised beta 2-microglobulin levels, while patients with Hodgkin's disease rarely show such increases in circulating beta 2-microglobulin.


Reference interval for maternal serum alpha-fetoprotein. Maternal serum alpha-fetoprotein (AFP) concentration was measured in 237 samples taken throughout the pregnancies of 28 normal pregnant women who all gave birth to normal infants. The patterns from these 28 individuals curves were compared graphically with an earlier published non-parametric 90% reference interval based upon single serum samples from other normal, pregnant women. The two "reference areas" were found to be quite similar. Maternal serum AFP values before and after induced abortion in a case of acrania and spina bifida demonstrated the value of serial maternal serum AFP quantitations in cases of suspected neural tube defects (NTD).


Comparison of alphafetoprotein and beta-trace protein in the antenatal diagnosis of anencephaly and spina bifida. Alphafetoprotein (AFP) and beta-trace protein (BTP) concentrations have been measured in amniotic fluids taken from 19 pregnancies where the outcome was an infant with a neural tube defect and from 19 matched controls. There was no significant difference in the mean BTP values for the pathological samples when compared with the controls, and  there were no striking differences in the values within individual matched pairs. On the other hand, the mean AFP concentration in the abnormal amniotic fluids was approximately 15 times the mean value found in the controls. Differences within individual pairs were particularly noticeable early in pregnancy. It is concluded that BTP is of no use for the early antenatal diagnosis of anencephaly and spina bifida.


["Trypanosoma brucei gambiense", antigenic study. III. Hemagglutination reaction with specific antigenic fractions]. The tannic acid hemagglutination method was applied to experimentally infected rodents sera. The antigens were Trypanosoma gambiense fractions separated by gel filtration on Sephadex G 200. Two fractions, at least, showed specific activity.


Surface immunoglobulins of lymphocytes in mouse plasmacytoma. IV. Evidence for the persistence of the effect of plasmacytoma-RNA on the surface immunoglobulins of normal lymphocytes in vivo and in vitro. The previous findings were confirmed that RNA extracted from murine plasmacytoma alters the character of the lymphocyte surface immunoglobulins (SIg) to express the idiotypic specificity of the Ig of the plasmacytoma from which the RNA was derived (cell conversion). RNA extracted from spleens of plasmacytoma-RNA-injected BALB/c mice also had convering activity, and if injected into other mice, caused the appearance of RNA active in cell conversion in spleens of the second set of mice. This activity was lost only after two additional transfers. When splenic cells from animals 1 hr after injection with RNA extracted from MOPC 300, LPC-1 or MOPC 104E, were cultured for 7 days, the proportion of cells with the SIg specific for these tumours increased. The cell-converting activity of the RNA extracted from the cultured cells after 7 days incubation ('7-day' RNA) was higher than that of RNA extracted from cells after 1 hr incubation ('immediate' RNA). 'Seven-day' RNA could be used for sequential transfers without marked loss of activity in cell conversion for at least five transfers. The repetitive transferability of this phenomenon by the injection of plasmacytoma-RNA suggests the possibility of RNA replication in the recipient cells.


Antiarrhythmic effect of Mercurascan in patients after surgical treatment of ischaemic heart disease. Premature ventricular contractions are relatively often encountered in patients after surgical treatment of ischaemic heart disease. Mercurascan (MSC), given at 0.05 mg/kg of body weight, favourably affected both the incidence and duration of arrhythmia. Unlike other antiarrhythmics, MSC did not affect the heart rate nor blood pressure. Postoperative treatment of ventricular arrhythmia with MSC is a promising procedure and merits further clinical and experimental investigation.


Centromeric and telomeric staining regions in the chromosomes of cattle (Bos taurus). Two new types of staining regions exist aftet treatment with a modified version of the ammoniacal-silver technique (Howell et al., 1975), in addition to findings by many other researchers working on the chromosomal complement of Bos taurus. Densely stained regions appear to be associated with the centromeres of many of the 29 pairs of acrocentric chromosomes, but there are also 4-6 of the same acrocentric chromosomes bearing telomeric staining regions (TSRs) at the ends of their long arms. In many metaphases examined, actual physical connectives were observed between both types of these ammoniacal-silver staining regions.


Speculations on the relevance of developmental psychology to paediatrics. Many paediatricians are now interested in applying the concepts, theories and methods of developmental psychology to clinical practice. The new understanding so obtained should enable abnormalities of clinical importance to be detected earlier; it should also provide means for evaluating the outcome of some disease processes and medical procedures more fully than was hitherto possible. The genesis of the nurturing failure or deficiency which underlies many paediatric problems becomes explicable as the nature of the maternal bond is unravelled. New ways may thus be opened for the more effective  prevention and treatment of child abuse, neglect and deprivation. Developmental psychology also promises to provide for the first time a framework on which child-rearing methods which will positively promote healthy psychological development can be based. However, some caution and critical interpretation, as well as much collaborative investigation, will be needed before the full relevance of developmental psychology to paediatrics is established with certainty.


Relationship between background activity and subclinical seizure pattern. Open- and closed-eyed EEG records were made in frontal, temporal and occipital regions, in order to examine the time relationships between various of their characteristics, in connection with the appearance of subclinical seizure patterns. 1. In the range of alpha frequency, both with open and closed eyes, the standard deviation of frequency over all brain areas decreased both before and after subclinical seizure patterns as compared to periods without pattern, and the number of temporal waves was reduced. Frontally, the mean amplitude was increased only in open-eyed records. The time relationship of the appearance of alpha maxima differed over all regions, but was closest in frontal records in both open- and closed-eyed records, and independent of the pathological pattern. In closed-eyed frontal records, a changeover in the time lead between the hemispheres could be observed, with one side leading during the period preceding the pattern, and the other after the pattern. 2. In the range of theta frequency, occipital open-eyed records showed slowing of the frequency both before and after the subclinical seizure pattern, with an increase in the standard deviation. In closed-eyed records made in the period preceding the pattern, an increase of the mean amplitude was observed frontally and occipitally, as well as an increase of the standard deviation of the amplitude in all regions. Slowing of the theta frequency was encountered in the frontal regions. Differences in the periods preceding and following subclinical seizures were to be seen only in the theta range.


Electrocorticography of waves associated with eye movements in man during wakefulness. Waves associated with horizontal saccadic eye movements were recorded from cortical occipital areas through multilead intracerebral electrodes implanted for a few days in 6 drug-resistant epileptic patients in order to localize epileptogenic foci. The waves were studied statistically when the EEG activity was not disturbed by interictal discharges. Cortical occipital waves related to free eye movements while scanning complex material started at the end of the eye movement, later in fact than the corresponding scalp parieto-occipital phenomena. The amplitude of the cortical waves increased with the complexity of the external field (contrasts having no marked effect) and was reduced in an unpatterned field and in dim light. Waves persisted in darkness. Their amplitude was independent of the size of the eye movement. The latency of the waves, calculated from the onset of eye movement, increased with the size of the movement, the complexity of the external field and also in darkness. Imposed eye movements induced cortical waves of larger amplitude and shorter latency and anticipatory potential changes beginning before onset of the eye movement. Waves related to eye movements differed from blinks and from responses to several types of light stimulation more strikingly in cortical records than in those from the scalp. These findings are discussed in relation to lambda waves, activation waves and eye movement potentials, expectancy waves and premotor potentials, and to the phenomena of perceptual blanks preceding normal perception after fixation of gaze. The importance of the subject's attention is emphasized.


Modulating systems of hippocampal EEG.

Modulating systems of hippocampal EEG. Hippocampal EEG changes in response to stimulation of several mesodiencephalic inputs (mesodiencephalic reticular formation, MRF; medial and lateral hypothalamus, MH and LH; medial septum, MS) were quantitatively investigated by means of spectral analysis in cats with chronically indwelling electrodes. The rhythmic activity of the hippocampus, in the theta frequency range, increased as a function of stimulus strength wherever the stimulation took place in MRF, LH and MH. Characteristic response patterns were found for each input area. These differed as regards the occurrence of shifts in theta peak frequency and the decrease in one or more theta frequency components below baseline levels at high stimulation voltage. Desynchronization (EEG flat spectrum) did not take place at low stimulation strength of the MRF, LH and MH except in a few cases, where the electrodes were placed in the neighbourhood of fibre tracts (fornix and medial forebrain bundle). Thus evidence for the existence of "synchronizing" and "desynchronizing" systems was not found. A model of the origin of the hippocampal EEG is discussed in the light of the results, by means of which several hypotheses concerning the mechanisms of desynchronization are considered.


Thalamic evoked potentials to somatosensory stimulation in man. Thalamic somatosensory evoked potentials (ThSEPs) were recorded by averaging technique from various thalamic structures during 59 stereotactic operations. From 372 records, response patterns and latency characteristics were analysed in relation to the intrathalamic localization. The findings can be summarized as follows. In N. ventro-caudalis (VC) and ventro caudalis parvocellularis (Vcpc) ThSEPs showed the most definite (and exclusively contralateral) responses characterized by a single positive (P1) deflection. The latency was shortest in VC (mean value, 17.5 msec) and in Vcpc (15.6 msec). Responses from N. centrum medianum (CM), parafascicularis (Pf) and limitans (Lim) were composed of early P1-N1 and of later P2-N2 components. The P1 latency was relatively consistent, with a mean value of 28.2 msec. Pulvinar responses showed a pattern similar to CM, with a mean P1 latency at 30.5 msec. Responses of N. dorsalis medialis (DM) were small, variable and longest, with a mean P1 latency of 54.2 msec. To ipsilateral stimulation, CM, Pul, DM and N. ventro-lateralis (VL) showed comparable wave forms. The possible role of the CM-Pf-Lim complex and of Pulvinar in the "extra-lemniscal" sensory system was considered. The usefulness of ThSEP recording to identify electrode locations in the thalamus is thus confirmed.


Epileptic discharges triggered by blinking and eye closure. Two children manifested EEG discharges which were triggered by blinking and/or eye closure in light and in darkness. The first case demonstrated single focal posterior discharges with eye blinks and repetitive focal posterior sharp wave discharges with eye closures. No photoparoxysmal responses were obtainable. Case 2 manifested generalized epileptic discharges triggered by eye closure which were sometimes associated with clinical seizures. In addition she showed photoconvulsive responses. Since the discharges triggered by eye blink and by eye closure occurred in both bright light and in darkness it is suggested that sensory afferents arising from contraction of the orbicularis oculi muscles may have been the epileptic trigger in Case 1 and could have contributed to the epileptic triggering mechanism in Case 2.


[Maturation of somesthetic evoked potentials in man]. Somaesthetic evoked potentials (SEPs) have been recorded on the parietal scalp of 375 normal and awake children to brief electrical stimulation (0.5 msec) applied to the contralateral median nerve at the wrist. Each SEP was obtained by summation of the responses to 150 stimuli delivered at a frequency of 1/sec. The population studied has been divided in five age groups, each one corresponding to a recognizable stage of maturation of the responses.


Comparison of Wiener filtering and selective averaging of evoked potentials. The application of the Wiener filter to the estimation of evoked potentials is criticized, and this method is compared with an a posteriori selective averaging method. It is shown that Wiener filtering may cause information loss for certain types of evoked potentials, since the transient evoked response components of the brain are of damped oscillatory character and are not stationary signals. The selective averaging method is briefly described and suggested to obtain consistent, dependable and more descriptive averaged evoked potentials (AEPs) of a brain structure for well-defined waking and sleep stages. The stated arguments are supported by a comparative representation of AEPs obtained from the cat inferior colliculus by means of Wiener filtering and selective averaging.


[High performance system for intracellular analysis]. A device is described, which allows intracellular microelectrode recording. It also has the possibility of injecting current without modifying the band pass, of eliminating artefacts, of controlling microelectrode resistance and capacity, measuring electrical membrane constants and, if need be, of unplugging the microelectrode.


Eectroencephalogram in "locked-in" syndrome. Twenty-three EEG studies in eight patients with "locked-in" syndrome were reviewed. The EEG was either normal or minimally slow in all except one patient. All showed reactivity to various stimuli. Photic stimulation elicited a photic driving response at one time or another in all the seven patients where it was attempted. Althouth the basic activity may be similar, it is the preservation of alertness and the presence of reactivity of the EEG which distinguishes the "locked-in" patients from those comatose due to an extensive brain stem lesion. In an unresponsive patient, an EEG resembling that of the waking state, although highly suggestive of an intrinsic brain stem insult, may also occur at times in association with diffuse cortical damage, secondary to prolonged cardio-pulmonary arrest.


Electroencephalographic changes during experimental hangover. The EEG was recorded in 27 subjects during hangover. Male healthy volunteers drank 1.75 g/kg body weight of ethanol in 3 h and the EEG was recorded 14-16 h later when the degree of hangover was highest. For control purposes a second EEG was recorded after a similar session when subjects drank water instead of ethanol. A third record was taken in normal laboratory conditions. T5-A1 and O1-A1 derivations were subjected to computer analysis from which spectral and frequency parameters were calculated. Visual analysis of the EEG during hangover showed a decrease and slowing of alpha activity and an increase in theta activity. Spectral analysis of the EEG gave a statistically significant increase in 7-8 c/sec activity during hangover. The EEG change could not be explained in terms of blood alcohol level, hypoglycaemia or acidosis. Also fatigue could be excluded as a cause of EEG change by means of "water controls". The conclusion is that the slowing of the EEG during hangover is caused by the depressant action of ethanol, or its metabolites, on cortical function.


[Contingent negative variation in  10-year-old children. Relations with the type of performance in a task of spatial localization]. CNV magnitude was studied in a task involving spatial localization and discrimination Subjects were 18 children 10 years of age. The CNV amplitude of half the subjects increased when the performance decreased and, in the other half, CNV amplitude decreased when the performance decreased. However, it seems that CNV magnitude increases as task difficulty increases but only above a minimal threshold varying with the subject. The results suggest that CNV amplitude cannot be related to a single psychological factor.


Effects upon the EEG of psychometric performance during deep dives in helium--oxygen atmosphere. A study of EEG modifications and psychometric performances was performed during two dives in hyperbarbic chambers with a helium--oxygen breathing mixture (pO2 maintained at 0.4 ATA). The depths attained were 500 m for the first experiment (Sagittaire II) and 610 m for the second (Physalie VI). Four subjects were studied. EEG modifications that could be considered as high pressure nervous syndrome began at 300 m. They were characterized by an increase of slow activity (particularly theta), depression of fast activity and transformation of the waking EEG into one resembling that of stage I sleep. The psychometric tests revealed a diminution of sensorimotor performance in all subjects (10--20%). Performance in intellectual tests varied according to subject and experiment, and did not fall more than 15% below normal values.


[Statistical analysis of the activity of pyramidal cells in the dorsal hippocampus of the rabbit]. The spontaneous activity of CA1 pyramidal cells was reduced with microelectrodes from the hippocampus of curarized adult rabbits under painless fixation. A statistical analysis of the data was made by a computer, using a program developed to process time interval series. 1. Various temporal patterns of discharge were observed. A classification into 5 distinct patterns could be disclosed, based on statistical parameters, particularly expectation density, joint interval histogram and interspike interval histogram. 2. The recorded neurones were distributed unequally in these groups, the distribution varying with sleep and wakefulness. 3. Some firing patterns prevailed during wakefulness and some during sleep, but all of them were observed in both states and no one was considered specific to one state. 4. The variability in neuronal discharge was estimated. "Stable" cells (60%) exhibited only one firing pattern. "Unstable" cells (40%) exhibited either two (82%) or three (18%) types of firing. 5. 65% of the cells recorded during waking and then during sleep shifted their firing pattern. The majority of the other units (35%), which kept the same pattern of discharge, were already stable during waking. Hence, they exhibited only one firing pattern and did not appear to be affected by sleep or waking.


Selective attention and the auditory vertex potential. I. Effects of stimulus delivery rate. In a selective attention task, twelve subjects received random sequences of 800 and 1500 c/sec tone pips in their right and left ears, respectively. They were instructed to attend to one channel (ear) of tones, to ignore the other, and to press a button whenever occasional "targets", tones of a slightly higher pitch, were detected in the attended ear. In separate experimental conditions the randomized interstimulus intervals (ISIs) were "short" (averaging 350 msec), "medium" (960 mes) and "long" (1920 msec). The N1 component of the auditory evoked potential (latency 80--130 msec) was found to be enlarged to all stimuli in an attended channel (both targets and non-targets) but only in the short ISI condition. Thus, a high "information load" appears to be a prerequisite for producing channel-selective enhancement of the N1 wave; this high load condition was also associated with the most accurate target detectability scores (d'). The pattern of attention-related effects on N1 was dissociated from the pattern displayed by the subsequent P3 wave (300--450 msec), substantiating the view that the two waves are related to different modes of selective attention.


Selective attention and the auditory vertex potential. Effects of signal intensity and masking noise. A randomized sequence of tone burst was delivered to subjects at short inter-stimulus intervals (mean ISI of 333 msec), with the tones originating from one to three spatially and frequency-specific channels. The subject's task was to count the tones in one of the three channels at a time, ignoring the other two, and press a button after each tenth tone. In different conditions, tones were given at high (60 dB SL) and low (20 dB SL) intensities and with or without a background white noise to mask the tones. The N1 component of the auditory vertex potential was found to be larger in response to attended-channel tones than in relation to unattended tones. This selective enhancement of N1 was minimal for loud tones presented without noise and increased markedly for the lower tone intensity and in noise-added conditions. The selectivity of attention was measured physiologically in this multichannel listening task was thus greater when tones were faint and/or different to detect.


Parameters of temporal recovery of the human auditory evoked potential. Auditory evoked potentials (AEPs) to tone pips at three monopolar scalters were systematically varied: tone intensity (3.0, 1.5 and 0.75 sec), and direction of attention. Interstimulus intervals were computed separately for the 9 different combinations of the three possible first prior intervals (intervals between the test stimulus and the stimulus immediately preceding it) and the three possible second prior intervals (intervals between the stimulus preceding the test stimulus and the stimulus prior to that). Our results show that temporal amplitude recovery of N1 and P2 can be based solely on the first prior interval had not effect on amplitude. Furthermore, they show that it is inadvisable to use combined N1-P2 amplitude measures since the two peaks appear to be governed by separate processes. Recovery for N1 was different from that of P2, N1 showed no intensity effects while P2 did, and N1 and P2 had different topographic distributions. Directing attention to the tones did not affect N1 or P2 amplitudes but caused a highly significant increase in both N1 and P2 latency. Attention to the tones also produced a frontal negative baseline shift following them.


The dimensionality of the human visual evoked scalp potential. A small number of processes can account for most of the evoked potentials activity in the two subjects studied. Principal components analysis indicates that six independent processes can account for approximately 97% of the variability in the data. Moreover, the factor analysis and plots of the factor coefficients yield indications that the times during which these principal factors are active agree quite well with the times at which the equipotential maps show some organized activity. The question of dipoles being the underlying cause of the observed activity is not answered by the factor analysis. The principal factors are not unique, but models which have a small number of parameters are more justifiable in light of the results of this study.


Fourteen and six c/sec positive bursts in comatose patients. Of ten patients with Reye's syndrome, there were five with stage II or III coma where EEGs revealed 14 c/sec positive bursts in a background of diffuse delta waves. Positive bursts disappeared upon EEG improvement in two survivors and when the EEG became nearly isoelectric in two other patients. Although 14 and 6 c/sec positive bursts are seen commonly during sleep in normal young persons, their occurence in association with diffuse delta waves in acutely ill, comatose patients has been rarely reported. It is not certain whether the present findings should be regarded as selective preservation of a type of sleep pattern or whether there are special factors that enhance positive bursts in stage II or III coma of Reye's syndrome.


Latencies of evoked potentials to flicker and to pattern speedily estimated by simultaneous method. This note describes a method for rapidly measuring the latency of a steady-state visual evoked potential (EP). Three sinewaves of different frequencies (F1, F2 and F3 c/sec) are summed, and the resulting waveform modulates either light intensity (for flicker stimulation) or spatial contrast (for pattern stimulation). Three evoked potentials of frequencies F1, F2 and F3 c/sec respectively are stimultaneously recorded by running three Fourier analysers in parallel. The outputs of the three analysers are presented in a single polar plot so as directly to display the three values of EP amplitude and phase. Practical methods for generating the pattern and flicker stimuli are described.


The contingent negative variation and psychological findings in chronic hepatic encephalopathy. Early diagnosis of chronic hepatic encephalopathy (CHE) in the latent stage before the appearance of clinical signs, should reduce both morbidity and mortality as deterioration is often preventable by treatment. Since existing diagnostic procedures are inadequate, we have investigated a test in which morphine is used as a provocative agent and any resulting change in cerebral function assessed by measurement of the CNV in conjuction with a psychological trail test. Twenty six patients were studied, 6 of whom had clinically overt CHE. A significant correlation (P less than 0.05) between the change in CNV amplitude with morphine and the initial CNV amplitude, consistent with the theoretical model of Tecce (1972), was found. However, the CNV and trail test results taken as a whole did not allow even those patients with overt CHE to be distinguished and we conclude that it is unlikely that differing degrees of latent CHE could be detected.


The stability of the sigma sleep spindle. The present study closely examined the distribution of sigma sleep spindle activity or six normal human males who slept undisturbed for approximately 8 h/night for 3 consecutive nights. Sigma spindle activity was monitored by an automatic spindle detector system which performed at 92.5% accuracy when judged against established visual criteria. Individuals differed markedly in total sigma spindle production across nights, but the failure to detect significant differences among nights and the large intra-class correlation suggests noteworthy inter-night stability of the sigma spindle.


Immunological comparisons of two synthetic human parathyroid hormone-(1-34) peptides. The immunological properties of human parathyroid hormone-)1-34) synthesized in accord with the sequence reported by Brewer et al. and the different sequence found by Niall et al. were compared with those of highly purified native human hormone-)1-84). Analyses were performed by radioimmunoassay using 7 anti-bovine parathyroid hormone antisera and bovine parathyroid hormone-)1-34) as tracer. Whereas human parathyroid hormone-(1-34) synthesized in accord with the sequence of Niall et al. was immunologically indistinguishable from native human hormone in all 7 assay systems, striking differences were seen between human parathyroid hormone-(1-34) synthesized in accord with the sequence of Brewer et al. and the native hormone. In none of the 7 assay systems did human parathyroid hormone-(1-34) synthesized in accord with the sequence of Brewer et al. give the same displacement slopes that the reference preparation gave. The observation that immunologic probes easily discriminate between the two different human parathyroid hormone-(1-34) peptides suggests that similar immunologic approaches will be of value in exploring several important issues, particularly those relating to the sequence and conformational properties of human parathyroid hormone and its synthetic peptides and to the question of the existence of isohormonal forms.


Antigenic determinants of murein lipoprotein and its exposure at the surface of Enterobacteriaceae. Murein lipoprotein from the outer membrane of Escherichia coli could be fixed to erythrocytes without pretreatment of the erythrocytes. Passive hemagglutination or immune hemolysis could thus be used as sensitive assays to determine antibodies against lipoprotein. In rabbit antisera prepared against whole cells of E. coli, Salmonella, Arizona, and Shigella antibodies against lipoprotein were present. The respective titers were lowest in encapsulated smooth strains and highest in rough mutants. Antisera against deep rough mutants showed even higher anti-lipoprotein titers than anti-R-lipopolysaccharide titers. Correspondingly,absorption of lipoprotein antibodies with enterobacterial strains was most pronounced with deep rough mutants and lowest with smooth strains. Lipoprotein becomes increasingly an immunogen as well as an antigen the more sugar residues are missing in the lipolysaccharide on the cell surface. In wild-type cells lipoprotein is buried in the outer membrane; its exposure in mutant cells is related to defects at the cell surface.


Total and free triiodothyronine and thyroid-binding globulin concentration in elderly human persons. Total thyroxine (TT4) and triiodothyronine (TT3) were found to be low in healthy elderly subjects with a preferential decrease of triiodothyronine. In order to determine the importance of these findings 22 healthy elderly subjects were examined. Free triiodothyronine (FT3), thyroid binding globulin (TBG) concentration and basal thyroid stimulating hormone (TSH) were measured by radioimmunoassay. Liver enzymes, cholesterol and total protein concentration were also assayed. TBG was significantly increased compared to a middle-aged group and did not correlate with TT4, TT3 and TSH. Basal TSH values were in the normal range and could be detected in all the elderly subjects in contrast to undetectable values in 40% of the younger subjects. FT3 determined directly did not correlate with the values calculated according to the law of mass action. According to the FT3 values the elderly subjects could be subdivided into three groups independent of their TT4, TT3, TBG and TSH values. FT3 was undetectable in one group, in the low normal to normal range in another and elevated in the third group. Our results suggest that 1) there is no correlation between TT4, TT3, elevated TBG and FT3 determined directly or by calculation, 2) basal TSH values seem to indicate possible hypothyroidism in elderly persons which is correlated with elevated cholesterol levels and 3) FT3 measured directly subdivides this metabolic state into three groups possibly depending on the intracellular concentration of T4.


Distribution of serum zinc between albumin and alpha2-macroglobulin in patients with decompensated hepatic cirrhosis. Lower concentrations of total serum zinc (540 +/- 111 mug/1, mean +/- SEM), and of albumin-bound serum zinc (295 +/- 113 mug/1) and a higher concentration of alpha2-macroglobulin-bound zinc (245 +/- 69 mug/1) were found in 25 patients with decompensated hepatic cirrhosis, compared to 28 healthy subjects (835 +/- 91; 679 +/- 83; 156 +/- 27 mug/1 respectively). Levels of total and albumin-bound zinc were significantly and positively correlated with serum albumin levels. Higher levels of alpha2-macroglobulin-bound zinc were associated with higher levels of alpha2-macroglobulin in these patients (2.8 +/- 0.8 g/1) compared to normals (2.3 +/- 0.6). Hence, not only do decompensated cirrhotics exhibit a lower serum zinc level but a greater proportion of this zinc is associated with the tightly bound, and presumably metabolically more inert, serum fraction. This situation exaggerates the zinc deficiency state of the severe cirrhotic.


A note on the method of retrograde transport of horseradish peroxidase as a tool in studies of afferent cerebellar connections, particularly those from the inferior olive; with comments on the orthograde transport in Purkinje cell axons. 1. Injections of horseradish peroxidase (HRP) suspension were made in the cerebellar cortex of cats (most often the paramedian lobule). The staining of the cerebellar cortex and the ensuing labeling of neurons in the inferior olive were studied in experiments with variations of concentration of HRP, amounts of fluid injected, survival time and age of the animals. Light microscopical studies were supplemented with electron microscopical observations. The folded cerebellar cortex offers particular difficulties with regard to obtaining a predictable extent of stained tissue, and the spreading of the fluid within the cortex shows great variations even with the same amounts and concentrations of HRP suspension. Diffusion of fluid appears to occur most easily within the molecular layer. Often there are unstained parts of folia between stained parts. Staining of the cortex is barely visible after 7 days, but appreciable shrinking of the stained area does not appear to occur until after 4 days. 2. The first signs of labeling of olivary neurons are seen after 5-10 hours, after 7 days there are no labeled cells. The rate of retrograde transport in olivocerebellar fibers is calculated to be between 50 and 100 mm/day. Labeling of cells appears to require staining of the molecular layer of its projection areas in the cerebellum. 3. For studies of the olivocerebellar projection survival times of 2-3 days and injections of 0.5 mul of a 50% HRP suspension seem in general to be well suited. Best results are obtained with animals weighing 1-3 kg. There is a clearcut correlation between the site of staining of the cortex of a particular part of the cerebellum and the site(s) and extension of olivary area(s) containing labeled cells. 4. Anterograde transport in axons of Purkinje cells has been observed. Electron microscopically the axons of these fibers contain HRP labeled tubules and vesicles as do their terminal boutons in the nuclei. 5. In cases where the injected fluid has spread to the cerebellar nuclei, localized parts contain neurons which are labeled as are the cells in the injected cerebellar cortex.


beta2-Microglobulins: isolation, properties, and distribution. beta2-Microglobulin is structurally related to immunoglobulin domains and is identical to the light chain of histocompatibility (HL-A) antigens. Similar to free light chains of immunoglobulins, beta2-microglobulin is most easily isolated from urine. We have previously purified human beta2-microglobulin from urine of patients with renal tubular resorption defects. Corresponding proteins have now been obtained from urine of rabbits and guinea pigs treated with sodium chromate. Sequence studies have established that the rabbit protein is rabbit beta2-microglobulin. The guinea pig protein closely resembles the human and rabbit beta2-microglobulins in amino acid composition, charge, molecular size, and also in the presence of an apparently analogous disulfide loop. These findings indicate that this protein is the guinea pig homologue of beta2-microglobulin. Physical-chemical studies suggest that human beta2-microglobulin and isolated immunoglobulin domains are similar not only in amino acid sequence but also in three-dimensional structure. Both types of molecules are compact and globular in shape and apparently contain beta-pleated sheet conformation. beta2-Microglobulin is present in free form in various body fluids and as a subunit of histocompatibility antigens on cell surfaces. Current estimates suggest that the number of beta2-microglobulin molecules on cell surfaces is higher than the number of histocompatibility (HL-A) antigens. Accordingly, beta2-microglobulin is possibly a subunit of additional cellular antigens or receptors.


Structure and significance of beta2-microglobulin. beta2-Microglobulin shares many structural features with the homology regions of the immunoglobulins. Particularly significant is the fact that its amino acid sequence is homologous to the sequences of the constant regions of both classes of light chains (kappa and lambda) and to the constant homology regions of at least three classes (gamma, mu and epsilon) of heavy chains, especially the carboxyl-terminal regions Cgamma3 Cmu4 and Cepsilon4. Molecules similar to human beta2-microglobulin have been found in other vertebrate species. The properties of beta2-microglobulin suggest that the gene for this protein may have evolved from a precursor gene that by duplication gave rise to immunoglobulin light and heavy chains. Furthermore, the observation that beta2-microglobulin is synthesized by and appears on the surfaces of a variety of cell types, including nonlymphoid cells, suggests that the concepts derived from analysis of the immune system may be applicable to other areas of cell biology. In particular, the close association of this immunoglobulin-like molecule with the histocompatibility antigens has a number of implications for the origin, structure, and function of these as well as other cell surface glycoproteins.


Some biological properties of beta2-microglobulin and its antibody. beta2-Microglobulin on the surface of lymphocytes exists in two molecular forms, first as part of the HL-A antigen complex, and second as a free molecule unbound to other membrane macromolecules. Quantitative determinations of the numbers of molecules of beta2-microglobulin per lymphocyte when compared to published values for the numbers of HL-A molecules per cell are consistent with an excess of beta2-microglobulin compared to HL-A antigens on the cell surface. Turnover studies of beta2-microglobulin from the lymphocyte surface indicated that both beta2-microglobulin and HL-A components are metabolized at similar rates. beta2-Microglobulin appears to be released in the free form. HL-A antigens, if released from the cell surface, appear to be released unbound to beta2-microglobulin. The effects of anti-beta2-microglobulin antibody on lymphocyte activation, namely on the mixed lymphocyte reaction and on antigen induced proliferative response, were studied. Anti-beta2-microglobulin completely inhibited the mixed lymphocyte reaction and the antigen induced proliferative response.


beta2-Microglobulin: a reevaluation.

beta2-Microglobulin: a reevaluation. Rabbit antiserum to human beta2-microglobulin was used to inhibit the proliferative response of lymphocytes in a variety of in vitro assays and to block the effector phase of the cell mediated lympholysis reaction. The antiserum was able to inhibit both of these reactions; it is not clear whether the cytotoxic reaction that we are studying in a xenogeneic human-rabbit system is based on phytohemagglutinin dependent cytotoxicity or on specific recognition of target cells by receptors on the effector cells, or most likely on a combination of both cytotoxic mechanisms. Whereas the possibility that beta2-microglobulin may be associated with receptors on the thymus-derived lymphocyte surface is considered, it is also pointed out that the effect of the antiserum may be based on other mechanisms of perturbing the membrane so as to inhibit these responses.


[The current role of palliative operations in the surgical treatment of tetralogy of Fallot. A report on 172 operated cases (author's transl)]. 172 consecutive palliative operations in patients with Fallot's tetralogy are presented. 19% of the patients were aged less than 6 and 45% less than 12 months. The overall mortality was 5.8%, but has been recently reduced with the improvement of anaesthesiologic and technical experience. In fact, during the last two years, 69 Waterston shunts (33 under 12, and 17 under 6 months of life) were performed with no deaths. On the grounds of this experience, indications of the different types of systemic to-pulmonary artery shunts are discussed, as well as problems related to the choice between one-stage and two-stages surgical correction. The authors' opinion is that palliative procedures still play an important role in the treatment of Fallot's tetralogy, although the future approach may well be early total correction. In their experience, the last approach is restricted to children over 18-24 months of life with favorable anatomy, and to all patients over 4 years of age.


The effects of portacaval shunting and portacaval transposition on serun IgG levels in the rat. In rats subjected to end-to-side portacaval shunt and to portacaval transposition, serum IgG levels rose progressively by approximately 100% over a 5-week period. During the same period, sham-operated control animals showed only the increase expected with age. Rats with a portacaval shunt showed a greater fall in body weight and liver weight than did those with a portacaval transposition, and also showed a greater fall in levels of liver-synthesized proteins. Serum enzyme levels were markedly elevated during the first 48 hr after portacaval shunting, whereas after portacaval transposition the elevation was very small. Over the following 5 weeks enzyme elevation continued to be margiallly greater in the portacavallly shunted animals. Because IgG levels rose to a similar degree in both groups of animals, the prsent results support the hypothesis that hypergammaglobulinemia is due to the shunting of antigen-rich portal blood past the reticuloendothelial cells of the liver, and the hepatocellular damage does not play a major role in this process. The etiology of hypergammaglobulinemia in chronic liver disease in man may be similar.


Diagnostic tissue microbiology methods.

Diagnostic tissue microbiology methods. Tissue specimens, when processed properly, may yield important microbiologic information. Various techniques have evolved for specimen collection and transport, processing of tissue specimens, and demonstration of organisms by staining histologic material that enhance the identification of infectious agents. Careful attention to these details may yield diagnostic information that might otherwise be missed.


The occurrence of antibodies in rabbit anti-pneumococcus antisera directed against calcium dependent and independent determinants. Two type specific acidic pol-saccharides from Pneumococci (type III and type VIII) have been used to prepare immuno-adsorbents. The polysaccharides were coupled to Sepharose 4b via a 1,6 hexanediamine spacer and used to isolate two populations of type specific antibodies from rabbit anti-pneumococcus antisera. One population requires calcium to affect interaction with the homologous polysaccharide, while the other population has no such requirement. The isolated antibodies are of the lgG class and are fully active in precipitin assays.


Effect of carbamylation of lysine epsilon-amino groups on the activity of blood group specific glycoproteins. Carbamylation of epsilon-amino groups of lysine of human blood group MM glycoprotein, some of its precursors and the blood group A B antigens gave products with 41% - 91% epsilon-amino group substitution. Even the most extensive carbamylation led to only marginal changes in the circular dichroic (CD) spectra of these substances and none in sedimentation coefficients studied. Nevertheless, carbamylation resulted in either increased or unchanged or decreased inhibitory activity of all blood group antigens tested depending solely on the source of the hemagglutinin used. Carbamylation of epsilon-amino groups of these blood group glycoproteins therefore leads to minor conformational changes, not involved with the primary blood group specificity, which is recognized by a large proportion but not by all corresponding antibodies and lectins.


Incompatibility between C3b and B of guinea-pig and man and its influence on the titration of the alternative pathway factors D and B in these two species. The incubation of the proteins C3b, B, D and C3 induces enzymatic activation of B and of C3. This system of purified components was applied to the titration of the guinea-pig and human factors D and B in various homologous and heterologous reaction mixtures. On the basis of functional as well as of immunological tests, the D enzymes of both species were found to be related proteins. First, titrations of factor B revealed that both D enzymes activate homologous and heterologous B with comparable efficiency. Secondly, a rabbit anti-D antiserum raised against guinea-pig D was found to form a precipitation line of partial identity with guinea-pig and human D. In contrast to the compatibility between heterologous D and B, heterologous combinations of C3b and B were found to be incompatible. This incompatibility  was indicated by low titres of D in the presence of heterologous C3b and B; in contrast, in the presence of homologous C3b and B, the titres of D were up to one hundred-fold higher. The reason for this effect was found to reside in inefficient complex formation between heterologous C3b and B. Therefore titrations of D and B should be only performed in the presence of homologous C3b and B.


Single cell studies on the antibody-forming potential of fractionated, hapten-specific B lymphocytes. This study addresses itself to the problem of antibody formation in vitro by mouse splenic B lymphocytes enriched for reactivity to the hapten NIP by the hapten-gelatine binding and melting technique of Haas and Layton (1975). Small numbers of NIP-gelatine-bound B cells were placed in microcultures either by bulk dispensing of dilute cell suspensions, or by micromanipulation under direct microscopic visualization. Antibody formation was induced by the T cell-independent hapten-protein conjugate NIP-polymierized flagellin, using 10(4) thymus cells per microlitre as 'filler' cells. The frequency of precursors of NIP-specific antibody-forming cells among bound cells was about 2-2 X 10(-2) (one cell in forty-five) by both statistical and direct evaluation, after adjustment for a background frequency of 6-10 X 10(-8) precursors in the thymus filler cells. Single clones commenced antibody secretion asynchronously, as shown by the fact that the incidence of positive cultures continued to rise over the whole three days of culture, and that very small clones  of one to four plaque-forming cells (PFC) were still found on day 3. The mean PFC number per positive culture rose from 1-2 at day 1 to 4-7 at day 2 and about 20 at day 3.


Influence of reticuloendothelial blockade on the induction of tolerance and immunity by polysaccharides. The intensity of reticuloendothelial blockade by carrageenan, silica and ethyl stearate was measured and its effect studied on the susceptibility to tolerance induction by polysaccharide antigens in mice. The most intense RE depression reduced the tolerance threshold dose of levan only by 3-fold and that of dextran B512 not at all. The genetic resistance of BALB/c mice to tolerization with the alpha1-3 glucose epitope of dextran B1355 was not overcome by carrageenan blockade which did however, render them normally susceptible to tolerance induction by human gamma-globulin. PFC responses to immunization by the polysaccharides were diminished by blockade, relative to its intensity and to the antigen itself. These and other data suggest that severe RE blockade (a) can promote tolerance by suppressing the active role(s) of the macrophages in immune induction rather than by sustaining circulating antigen, and (b) depresses responses to thymus-independent antigens, probably by an immunosuppressive influence mediated by damaged macrophages.


Quantitative and thermodynamic measurements on I and i antigens of human red blood cells. Different homogeneous IgM cold agglutinins (two anti-I, two anti-i and one anti-Il cross-reacting antibodies) have been used to determine the antigen site densities of adult I and i erythrocytes and of cord red cells. The equilibirium constants and the thermodynamic constants of these reactions have been determined. The two anti-I antibodies, which did not combine with i or cord red blood cells, recognized two different determinants on I red cells. The antigen density of the I Fla. receptor was 120,000 sites per erythrocyte and the standard enthalpy change (-deltaH degrees) of the reaction was 18 to 25 kcal/mole. The antigen density of the I Loi. determinant varied according to the red cells tested and the enthalpy change (-deltaH degrees) of these reactions was 50-65 kcal/mole. The i and cord erythrocytes reactive structures were more heterogeneous than those present on I erythrocytes. The equilibrium constants rapidly decreased as  the temperature rose and the standard enthalpy changes (-deltaH degrees) ranged from 50 to 90 kcal/mole. Two types of i determinants were observed; one of the anti-i antibodies reacted mainly with an i component present on cord erythrocytes, the other antibody reacted with a different i component present on i adult red-cells. The determinants, recognized by the cross -reacting antibody on I red cells, differed fromthose on i or cord red cells in equilibrium constant, thermodynamic constants, index of heterogeneity of the reaction and in their sensitivity to formalin treatment.


Isoprinosine augmentation of phytohemagglutinin-induced lymphocyte proliferation. The need for agents designed to modify immune response in the treatment of patients with viral infection, immunodeficiency, or cancer prompted the present study on the mechanisms of action of isoprinosine, a compound developed for antiviral use and whose therapeutic activity may involve the immune system. The effect of isoprinosine on in vitro proliferation of human peripheral blood lymphocytes stimulated by phytohemagglutinin (PHA) and on lymphocyte levels of cyclic adenosine 3',5'-monophosphate and cyclic guanosine 3',5'-monophosphate was analyzed. Over a concentration range from 0.2 to 250 mug/ml, isoprinosine augmented PHA-induced proliferation; maximal stimulation was observed between 25 to 50 mug/ml. Isoprinosine in the absence of PHA had no effect on proliferation. The relative lack of effect of isoprinosine during a 90-min exposure and the lack of effect on lymphocyte cyclic nucleotide levels indicate that isoprinosine potentiates the PHA response by a mechanism different than a number of hormonal agents and such immunopotentiators as levamisole, polyadenylic-acid, and endotoxin. Further evaluation of isoprinosine as an immunopotentiator is indicated.


Antigenicity of the M proteins of group A hemolytic streptococci: further evidence for shared determinants among serotypes. Shared antigenic determinants between M proteins of group A streptococci appear to be widespread among serotypes. This is demonstrated by the ability of purified M proteins to absorb opsonic antibody from a variety of heterologous antisera prepared against whole cells or purified M proteins. This absorption procedure has the capacity to separate passive mouse protecting and passive hemagglutinating antibodies from opsonic antibodies measured in vitro. When whole cells or M proteins are used as heterologous absorbents, immunoglobulin may be recovered from the cells or precipitates. The recovered antibody has most of the opsonic and some of the precipitating qualities of the original unabsorbed serum, but hemagglutinating titers are significantly lower. These data provide additional evidence that shared antigenicity among M proteins can be the result of common antigenic determinants. Arguments are presented that these cross-reactions are not the result of a nonspecific protein fraction associated with purified M proteins.


[A new instruction theory: possibility of a reverse flow of information from polypeptide sequences to RNA particularly in antibody synthesis, and the mechanisms of tolerance induction and immunosuppression (author's transl)]. A new instruction theory for antibody formation is presented. The reverse flow of information from the amino-acid sequences of small antigenic determinants to an antideterminant RNA (aRNA) seems feasible. Prerequisites are specific activating enzymes, tRNAs, ATP as well as some kind of membrane assembling the anticodons of tRNAs linearly, analogous to the linear primary structure of stretched polypeptides. Once synthesized, aRNA might be replicated, utilized as transfer factor and transcribed by means of Reverse Transcriptase into aDNA. Further steps would be the fusion of this aDNA with genetical performed DNA-molecules already coding for the basic strucures of different classes of immunoglobulins by means of a terminal deoxynucleotidyl-transferase. This could be a chromosomal or extrachromosomal integration. The second hypothesis concerns antigen-induced immunosuppression and the phenomenon of nonresponsiveness (tolerance). An overwhelming proteolysis might give rise to a degradation of antigens or receptor templates for antigenic determinants located on the surface of macrophages. On later exposure to a similar antigen proteolytic enzymes are already preformed abolishing rapidly antigenic information. The third hypothesis concerns antibody-induced immunosuppression and tolerance. Antideterminant information is integrated into the genome or established extra-chromosomally. The continuous presence of antibodies sets in motion a sequence of reactions causing an accumulation of all information intermediates including a complementary DNA strand to the aRNA. On exposure to the corresponding antigen aRNA is transcribed. However, translation might be inhibited by hybridisation with the complementary aDNA strand as well as specific RNA hydrolysis by RNase H. Concerning the immunogenity of antibodies, a proteolytical mechanism might also be possible. Taking this into account a tolerance could be suspended in the following way: 1. by influencing the overwhelming proteolytical degradation of antigenic determinants with simultaneous antigenic stimulation; 2. by substitution of aRNA to induce blocked antibody synthesis.


Investigation of the serum level of pregnancy-associated alpha2-globulin during contraceptive treatment by means of radial immunodiffusion method. The serum level of pregnancy-associated alpha2-globulin was studied in 70 healthy women taking three different types of oral contraceptives [40 Bisecurin: containing 1.0 mg of Ethynodiol diaceticum +0.05 mg Ethynilestradiol; 20 Infecundin: containing 2.5 mg of Norethynodrel +0.1 mg of Mestranol; 10 Cervicundin: containing of 0.5 mg Ethynodiol diaceticum]. Elevated amounts of serum level of pregnancy associated alpha2-globulin could be detected in 37 of the 40 women taking Bisecurin and in all 20 taking Infecundin. Cervicundin however, failed to induce any change in the serum protein pattern of the 10 subjects studied. Serum levels of pregnancy-associated alpha2-globulin were higher in the Infecundin-taking (53 mg %), than in the Bisecurin-taking group (29 mg %). This shows the oestrogen dependency of the pregnancy-associated alpha2-globulin and points to a possible implication of this globulin in the immunosuppressive processes.


Automatic gram-staining with the microstainer II. A comparison was made between the Microstainer II, an automatic staining machine, and the traditional, manual gram-staining method using clinical material and known organisms in a double blind study. Gram-reactions were in agreement with 98.4% of the organisms. The machine-stained microorganisms were generally found to be of the same or better quality than manually-stained organisms. Transfer of bacteria from slide to slide or smear to smear was not a significant problem. The Microstainer II would appear to be a useful addition to the large volume bacteriology laboratory.


Histochemical localization of estrogen induced sulfated glycoprotein in rabbit uterus. Histochemical localization of the estrogen-induced sulfated glycoproteins was made in the estrogen-treated rabbit uterus. Biochemical studies by a group of Endo et al, affirmed these particular glycoproteins were PAS-positive and metachromatic as stained with TB. No sign of digestion, however, has been detected in a series of tests with alpha-amylase, testicular hyaluronidase, streptomyces hyaluronidase, chondroitinase AC and chondroitinase ABC, and heparinase. The apical portions of the epithelial and glandular cells, obviously expanded by the estrogen treatment, display strong beta-metachromasia with TB (pH 4.0), saliva-resistant PAS-positive reactions, and also alcianophilia with AB (pH 2.5). These reactions are not reduced after the treatment with the enzymes above-mentioned. Meanwhile, in the stromal matrix, the same enzymes give an influence to diminish the reactions to various extent. Our results suggest that the estrogen-induced sulfated glycoprotein is definitely localized in the apical portions of the epithelial and glandular cells. The identity is emphasized between the substance that is elucidated in the histochemical sections and the sulfated glycoproteins that have been specified solely by means of biochemical assays.


The effect of graded 60 degrees C 1N nitric acid extraction and of deoxyribonuclease digestion on nuclear staining by metachrome mordant dye metal salt mixtures. We can divide metachrome mordant staining of nuclei after graded 60 degrees C 1 N nitric acid extraction into three groups. The Feulgen nucleal reaction and dilute cationic dye staining of nuclei are abolished in about 30 minutes. With one group of metachrome dyes nuclear staining is lost with acid exposures of one hour or less. In a second group nuclear staining is weakened by 30-60 minute extractions, but persists in recognizable grade for 4-6 hours. In the third group nuclear staining remains almost unimpaired for 4-6 hours. In the first group the nuclear staining seems clearly assignable to the nucleic acids and to DNA in particular. In the second group loss of part of the reactivity on short exposure indicates some participation of DNA in the control staining result, as well as participation of basic nucleoprotein. In the third group staining seems assignable largely to basic nucleoprotein. The five gallocyanin group dyes, all in group 1, all possess a dialkylamino group, probably functioning as an ammonium chloride.Hematoxylin, the flurone blacks and gallein all present an o-hydroxysemiquinone group which probably acts as a weak acid, in addition to the carboxyl group of gallein which gives the strongest staining of nuclei at the longest acid exposure. Deoxyribonuclease digestion (2 hours, 37 degrees C) separated sharply a class in which nuclear staining failed completely, a class in which nuclear staining was fully equal to that in the control preparations and an intermediate group in which slight, moderate, or severa impairment was present. Generally there was good agreement between the two methods of nucleic acid removal, despite the fixation difference. In each case, however, the extraction procedure was one worked out for the fixation on which it was used.


The activity of mitochondrial alpha-glycerophosphate dehydrogenase in the rat nephron following triiodo-L-thyronine treatment: a histochemical study. The effect of triiodo-L-thyronine (T3) treatment (15 mug/100 g body weight daily for 10 days) on the activity of mitochondrial alpha-glycerophosphate dehydrogenase (GPOX) in different nephron segments of the male rat was investigated by a histochemical staining method. The study showed marked segmental differences regarding the response to T3-treatment: 1. The first two proximal segments were unstained in the control rats and intensely stained following treatment. 2. The third proximal segments, the thick ascending limbs of Henle's loop and the distal convolted tubules showed a strong or moderate reaction in controls and a moderate increase after T3-treatment. 3. The high activity of collecting ducts in the cortex and outer zone of the medulla in controls was slightly increased by treatment. 4. Faintly reacting glomeruli and negative thin limbs of Henle's loop and collecting ducts in the inner medulla (papilla) were unaffected by T3-treatment. The results are discussed in relation to biochemical and physiological data.


On the histochemistry of N-acetyl-beta-D-glucosaminidase in rat central nervous system. The optimal conditions for histochemical demonstration of NAG activity in the cerebrum, diencephalon, midbrain, cerebellum, medulla oblangata, and spinal cord were studied in a series of 37 Wistar rats of either sex. The following more important results were obtained: Each CNS zone required a definite methodlogical approach. Optimal fixation for most structures was achieved after 2 h treatment with formol-calcium and subsequent immersion of tissue blocks in formol-calcium 0,88 M saccharose. By this fixation technique it was possible to preserve high enzyme activity and good tissue structure. Only for the large pyramidal cells of the cerebral cortex the method of Holt provided optimal fixation. Formol-calcium-saccharose mixture and pure 0,88 M saccharose produced the opposide osmotic effect on nervous tissue previously fixed with formol-calcium: the former induced tissue shrinkage, the latter edema. The use of hexazonium p-rosaniline coupler prompted preliminary alcohol treatment of sections and introduction of 0.1 M acetate buffer in the incubation solution. Acetate buffer concentrations lower than 0.2 M diminished the diffuse cytoplasmic coloration and permitted a clear-cut demonstration of the lysosomal reaction. Ample information on the distribution of NAG activity in the CNS was obtained by using fast garnet GBC coupler and 0.1 M citrate buffer. Manganese chloride in a 0.2 mM concentration activates the reaction. The distribution of NAG reaction product in the cells of the different sections of the CNS was studied. The distribution of NAG reaction product in the cells of the different sections of the CNS was studied. The neurons, glial cells, and blood vessels showed positive reaction. Strongest activity was reported for the neurons of the supraoptic and paraventricular necleus, the epithelial cells of the chorioid plexus, nucleus ruber of the mesencephalon, and the vascular wall pericytes.


Basis of microheterogeneity of myelin basic protein. The basic protein of bovine central nervous system myelin contains a single polypeptide chain of 170 amino acids. Multiple components of basic protein have been observed on disc gel electrophoresis and ion exchange chromatography at alkaline pH, but the basis of the microheterogeneity has not been established. In the present study myelin basic protein from bovine spinal cord was chromatographed on carboxymethylcellulose at pH 10.4 in glycine buffer/2 M urea. Three major peaks were obtained, identified as components 4, 5, and 6 in the oder of their elution from the column by a linear salt gradient. The amino acid compositions of tryptic peptides from components 4 and 6 were identical and the COOH-terminal sequence, Ala-Arg-Arg, was intact for all three components. Component 4 was found to differ from component 6 by partial phosphorylation of threonine 98 and serine 165. This modification was estimated to account for 50% of component 4. Component 5 differed from component 6 by partial deamidation of glutamine residues 103 and 147, which accounted for 80% of this component. These modified glutamine residues were also present in component 4 and constituted another 15% of this component. It was considered that component 6 was the native, unmodified species of basic protein and that component 4 differed by a net negative charge of 2, and component 5 by a net negative charge of.1 as a result of these modifications. The nonrandom nature of the modifications suggested the involvement of specific enzymes.


Role of a lipopolysaccharide gene for immunogenicity of the enterobacterial common antigen. It is known that only certain strains of the family of Enterobacteriaceae, notably rough (R) mutants with the type R1 or R4 core, evoked antibodies in high titers against the common enterobacterial antigen (CA) after immunization of rabbits with heated cell suspensions. The present investigation deals with genetic and immunochemical aspects of certain R1 and R4 mutants isolated from Escherichia coli 08 and various Shigella serotypes which, unexpectedly, do not induce CA antibody formation. Immunochemical and genetical (transduction and conjugation) experiments revealed that the rough phenotype of these special mutants was evoked by a mutation of pyrE-linked rfa gene, called rfaL, which is involved in translocation of O-specific polysaccharides onto the lipopolysaccharide core. The transduction of the defective rfaL, allele into appropriate rough recipients results in transductants which have simultaneously lost the ability to evoke CA antibodies. This finding suggests that a close connection exists between the function of the rfaL gene and the expression of CA immunogenicity in R1 and R4 mutants. One of the strains synthesized neither O-hapten nor CA, suggesting a mutation in a region equivalent to the rfe genes of Salmonella.


Structure and function of beta-blucosidases in Dictyostelium discoideum. There are two isozymes of beta-glucosidase in developing cells of Dictyostelium discoideum. A procedure for screening large numbers of clones for beta-glucosidase activity was utilized to obtain mutations which directly affect the activity. We recovered seven strains which lack both isozymes and four strains with residual activity in which enzymatic and physical properties of both isozymes are altered. Beta-Glucosidase appears to act as a block to selfing in macrocyst formation as shown by the fact that ssite mating type to form macrocyst-like structures. Immunological evidence utilizing antisera prepared against purified beta-glucosidase-1 demonstrates that most of the glycosidases in Dictyostelium discoideum share a common antigenic determinant which appears to be added post-translationally. The two isozymes of beta-glucosidase share common protein subunits but the antigenic determinant is either lacking or masked in beta-glucosidase-2. This may account for some of the enzymatic and physical differences between the two isozymes.


Ultrastructure of acetylcholine receptor clusters on cultured muscle fibers. The structure of regions with a high concentration of ACh receptors (clusters) on cultured skeletal muscle myotubes was examined by immunoperoxidase staining of bound alphaBT. The clusters did not appear to differ from the other regions except in their higher concentration of receptor.


Pleiotropic phenotype of colchicine-resistant CHO cells: cross-resistance and collateral sensitivity. Colchicine resistant (CHR) mutants of CHO cells with reduced permeability to colchicine display extensive cross-resistance to a number of apparently unrelated compounds including puromycin, daunomycin, emetine, ethidium bromide and gramicidin D. A positive correlation was observed between the level of cross-resistance and the relative hydrophobicity of these compounds. The mutants also showed increased (collateral) sensitivity to local anaesthetics (procaine, tetracaine, xylocaine and propanolol), steroid hormones (1-dehydrotestosterone, corticosterone and 5beta-pregnan-3,20-dione) and some Triton X compounds. In general, the degree of the pleiotropic response (cross-resistance or collateral sensitivity) correlated with the degree of colchicine resistance in mutant lines. These results are consistent with the pleiotropic phenotype being the result of the same mutation(s) which confer colchicine resistance and support a model for resistance in which the reduced permeability is assumed to be the result of an alteration in the modulation of the fluidity of the surface membrane.


Electron microscopy of interphase chromosomes in situ; binding of permanganate to chicken erythrocytes. From quantitative electron-microscope observations on the binding of permanganate to regions of erythrocytes and reticulocytes of known chemical composition, it is concluded that KMnO4, like phosphotungstic acid (PTA), binds preferentially to sites on proteins. Compared with PTA, KMnO4 binding exhibits less anomalous behaviour. The data support the hypothesis previously put forward that the 2 regions, or phases, in condensed chromatin differ in both molecular composition and concentration. The increase in binding to protein which occurs during nuclear haemolysis is interpreted in terms of protein-protein interaction in the chromatin of the intact cell.


Evaluation of a Proteus rettgeri O-serotyping system for epidemiological investigation. An evaluation of the serotyping system for Proteus rettgeri has been performed through its application to 561 human isolates of this species. Of these, 545 were typable and could be separated into 58 O serotypes. A group of isolates from one hospital, and clinically implicated in cross-infections, was found to belong the same O serotype, confirming the clinical findings. Isolates with more than one specificity of thermostable antigen, in most cases, were found to have combinations of such specificities restricted to combinations previously defined for the O-type strains. Furthermore, the use of pooled antisera was investigated and system of pooling was devised to retain the specificity of the system and to increase the facility of the typing procedure.


Defective histamine release in chronic urticaria. Histamine release from peripheral blood leukocytes challenged with anti-human IgE was studied in patients with chronic urticaria and nonatopic controls. 19 of 23 controls, but only 6 of 20 patients, released over 20% of the total available leukocyte histamine. The response to anti-IgE concentrations of 1.66, 0.33, 0.066, and 0.013 mug antibody N/ml was significantly lower in patients than in controls. Serum IgE levels were significantly higher in the patients but total histamine content of about 10(7) leukocytes was not. Deuterium oxide (D2O) greatly increased histamine release (in both groups), indicating that the anti-IgE interacted with the basophils of urticaria patients. Passive sensitization of leukocytes with biologically active IgE was achieved in both patients and control subjects whose cells responded to anti-IgE, but was not achieved in either patients or control subjects whose cells were nonresponsive to anti-IgE challenge. 125I-anti-IgE autoradiographic studies revealed no obvious quantitative abnormality in the amount of basophil-bound IgE in chronic urticaria patients. Ionophore stimulation of aliquots of the same leukocytes used for anti-IgE challenge demonstrated that the urticaria patients' basophils were capable of releasing normal amounts of histamine. Leukocyte cyclic AMP levels in the two groups were not significantly different either in base-line levels or in responsiveness to stimulation with isoproterenol. These data indicate that chronic urticaria patients have a (acquired?) defect in leukocyte histamine release that occurs after the anti-IgE-IgE interaction, but before the actual (second-stage) release process, and that is comparable to the phenomenon of desensitization.


The effect of immunotherapy on humoral and cellular responses in ragweed hayfever. The effect of specific immunotherapy on several in vitro responses to ragweed antigen E has been evaluated in 17 atopic patients with ragweed hayfever. The methods employed were leukocyte histamine release, measurement of specific IgE anti-ragweed antibody and specific IgG anti-ragweed antibody, lymphocyte proliferation, and the production of two lymphocyte mediators (migration inhibitory factor and mitogenic factor). The duration of treatment and symptom improvement were also recorded for comparison. Immunotherapy was associated with a decrease in leukocyte sensitivity for histamine release to ragweed antigen E in a majority of the patients. In addition, there was a significant decrease in IgE anti-ragweed antibody and a significant increase in IgG anti-ragweed antibody. Immunotherapy also resulted in a significant decrease in lymphocyte responsiveness to ragweed antigen E as measured by proliferation and the production of mediators. Symptomatic improvement was best correlated with the presence of IgG anti-ragweed antibody responses. The production of this antibody was also associated with a decrease in lymphocyte responsiveness. The results of this study indicate that specific immunotherapy in ragweed-sensitive patients induces alterations in immunologic reactivity to ragweed antigen in vitro. This response is antigen specific, includes elements of both humoral and cellular immunity, and may account for the clinical improvement that is often observed in patients who undergo this form of therapy.


The preparation, use, and evaluation of recorded material for teaching ultrasonics in a university Master of Science course in medical physics. The potential value of using recorded instructional material in a university Master's course module in medical ultrasonics is discussed. The procedure for preparation and evaluation of a program of slides with synchronized audio tape is reported. The evaluation indicates that the flexibility of this form of recorded material can make several unique contributions to the teaching program, in acquainting students with clinical problems, in simulating expensive equipment, and encouraging students to use the literature.


A phase I study of 5-azacytidine (NSC-102816). 5-Azacytidine was administered daily to 12 patients in a five-day schedule and to 15 patients in a weekly schedule as part of a phase I trial. The daily dose ranged from 50 mg/m2 to 158 mg/m2 and the weekly dose, from 200 mg/m2 to 633 mg/m2. The maximum total dose was 2000 mg in the daily schedule and 3775 mg in the weekly schedule. The major toxicity was gastrointestinal, with nausea and vomiting occurring in all patients in this study. Myelosuppression was less frequently encountered and appeared to be related to the increase in 5-azacytidine dose. Patients receiving 5-azacytidine in a weekly schedule of administration appeared to tolerate the drug better and to be more willing to continue their therapy.


Nigrothalamic projections in the monkey demonstrated by autoradiographic technics. In spite of repeated demonstrations by degeneration technics, nigrothalamic fibers have been regarded with some skepticism. Attempts were made to trace nigral efferent projections in the monkey by autoradiographic technics. Tritiated amino acids (L-leucine, L-lysine and L-proline), injected into portions of the substantia nigra (SN), labeled cells in four regions, designated as, (1) rostrolateral, (2) caudolateral, (3) rostromedial and (4) central. Rostrolateral nigral neurons transported radioactive label preferentially and abundantly to thalamic nuclei; localized isotope was found in parts of three thalamic nuclei, the medial part of the ventral lateral nucleus (VLm), the magnocellular part of the ventral anterior nucleus (VAmc), and the paralaminar part of the dorsomedial nucleus (DMpl)9 Lateral neurons in the caudal half of the SN transmitted radioactive label to the same thalamic nuclei as rostrolateral nigral neuron. Isotope transported to portions of the striatum was modest and localized. Radioactive label taken up by large cells in the caudal third of the SN was transported to portions of the striatum, but not to thalamic nuclei. Labeled nigral neurons in the medial two-thirds of the rostral half of the SN, and in the middle third of the central part of the SN, transported the label mainly to parts of the caudate nucleus and putamen. In these animals modest radioactive label was seen in VLm and VAmc, but not in other thalamic nuclei. There was no evidence that nigral neurons project to the subthalamic nucleus. No radioactive transport from nigral neurons was detected in the superior colliculus, the midbrain tegmentum, or the red nucleus, and none was transported to more caudal brain stem nuclei. Nigrothalamic fibers arise particularly from cells in rostral and lateral parts of the substantia nigra. While some cells in other parts of the nigra project to thalamic nuclei, these appear scattered and less numerous. Large cells in caudal parts of the SN do not project to thalamic nuclei. These observations confirm nigrothalamic projections to VLm and VAmc, and identify a new nigral projection to part of the dorsomedial nucleus of the thalamus (DMpl). No nigral efferent fibers project to any of the intralaminar thalamic nuclei.


Entry of peroxidase into neurons of the central and peripheral nervous systems from extracerebral and cerebral blood. Autonomic preganglionic, sensory, and lower motoneuron perikarya within the central nervous system, as well as cell bodies with axons projecting to the circumventricular organs, are retrogradely labeled with horseradish peroxidase (HRP) delivered to their axon terminals by cerebral and extracerebral blood. Subsequent to vascular injection of HRP into mice, blood-borne peroxidase passes across permeable vessels in muscle, ganglia, and in all circumventricular organs except for the subcommissural organ in which no leak could be discerned. Brain parenchyma adjacent to each of the permeable circumventricular organs quickly becomes inundated with the protein. By four to six hours post-injection, this extracellular HRP reaction product has disappeared, and by eight hours perikarya of specific hypothalamic nuclei contain HRP-positive granules indicative of the intra-axonal retrograde transport of the protein. Hypothalamic neurons so labeled are presumed to send axons to such circumventricular organs as the median eminence or neurohypophysis and include neurons of the magnocellular neurosecretory supraoptic and paraventricular nuclei, the accessory magnocellular nuclei, the parvicellular arcuate nucleus, and a band of periventricular cells extending rostrally into the medial preoptic area. Labeled somata are also adjacent to the organum vasculosum of the lamina terminalis and in the vertical limb of the nucleus of the diagonal band of Broca. No similarly labeled cell bodies were identified near the subfornical organ.


Effect of ragweed hyperimmune human gamma globulin on in vivo and in vitro parameters of ragweed sensitivity. The effect of ragweed hyperimmune human gamma globulin upon in vitro and in vivo parameters of ragweed sensitivity was examined. In a double-blind study, 40 ragweed sensitive patients were divided into 2 groups and received either ragweed hyperimmune human gamma globulin (treated) or normal pooled globulin (control). Parameters of ragweed sensitivity studied before and following injections of the gamma globulin included skin test and nasal provocation end point titration, specific IgE antibody as measured by the RAST, leukocyte histamine release, total serum ragweed antibody level, and total serum IgE. Changes in measured parameters varied in both groups of patients. In the "treated" patient group, 1 wk later nasal sensitivity decreased significantly, and there was a trend toward decreased histamine release from leukocytes. No discernable effect was noted upon the other parameters. Thus, with the dosage used, the parenterally administered hyperimmune gamma globulin did not influence most measurements of ragweed hypersensitivity. The concept of this therapeutic approach might warrant further investigation.


Alternaria IgG precipitins and adverse reactions. Late reactions consisting of fever, malaise, and swelling at the site, 4 to 6 hr after injections of Alternaria extract occurred in several patients receiving immunotherapy with Alternaria. These patients had in common serum IgG precipitins and exquisite leukocyte histamine release sensitivity to Alternaria. Such precipitins were 3 times more frequent in patients receiving Alternaria immunotherapy than a control group of patients receiving immunotherapy with other antigens. A prospective study revealed that 5 of 23 Alternaria-sensitive persons had precipitins before immunotherapy and another 6 developed precipitins during therapy. Only one of the 23 experienced a late Alternaria reaction. Thus, precipitins to Alternaria are common and do not seem to be the basis for the late reactions we observed. The finding of precipitins does not contraindicate immunotherapy.


Inhibition of antigen-induced histamine release by ouabain. The effect of ouabain, a specific sodium-potassium dependent adenosine triphosphatase (Na+-K+-ATPase) inhibitor, on antigen-induced histamine release was studied using guinea pig lung fragments sensitized in vitro with rabbit antibodies against bovine serum albumin. Histamine was assayed spectrofluorometrically. When sensitized tissue had been preincubated with ouabain (less than or equal to 1.0 x 10(-4) M) for 10 min prior to antigenic challenge, release of histamine was significantly inhibited (maximum 54%, p less than 0.001, N=9, paired t test). The most significant inhibition was obtained near the optimal concentration of antigen. The inhibition was dependent on the length of preincubation (less than or equal to 20 min), and was partially reversible upon washing the tissue removing the ouabain. Ouabain did not seem to prolong the duration of the histamine release process. Increase in potassium ion (less than or equal to 1.1 x 10(-2)M) inhibited the histamine release and had additive effects to ouabain action. Dibutyryl cyclic AMP (less than or equal to 5 x 10(-3) M), which could enhance the release, strongly antagonized the inhibition. Glucose removal from the medium did not abolish the ouabain effect. The results seem to indicate that immunologic release of histamine is under the influence of the membrane Na+-K+-ATPase activity.


Clinical and immunological studies of timothy antigen D immunotherapy. The response to preseasonal immunotherapy with aqueous grass extract, timothy antigen D, or water-soluble timothy (WST) in alginate was compared in patients sensitive to grass pollen. Injections of antigen D in alginate produced little evidence of clinical or immunologic response. Treatment with aqueous grass extract or WST in alginate, on the other hand, significantly reduced the seasonal rise in grass-specific IgE. Aqueous extract therapy was also associated with a decline in leukocyte sensitivity during the pollen season, while WST treatment produced the greatest rise in hemagglutinating antibodies.


Carcinoma of the vulva.

Carcinoma of the vulva. Carcinoma of the vulva often presents as a large ulcerating or fungating tumor with considerable destruction of the vulva and surrounding tissues. The area usually is very painful and foul smelling. Most such tumors are of the squamous-cell variety. Despite the degree of histologic differentiation, the lymph nodes are involved in more than 50% of the cases. The best therapeutic results are obtained with radical vulvectomy and groin node dissection "en bloc." Occasionally treatment should be modified in the geriatric patient because of associated medical problems. Various techniques are discussed. Since the disease is rare, best results are achieved by referral to a specialized treatment center.


Analytical electron microscopic study of mitochondrial inclusions in canine myocardial infarcts. An analytical electron microscopic study, utilizing scanning transmission electron microscopy and energy-dispersive x-ray spectroscopy, was made of two types of mitochondrial inclusions identified in canine myocardial infarcts. The data were obtained from thin sections of tissues that were fixed in aldehyde, osmicated and embedded in epoxy resin. Calcium peaks of variable intensity were detected in inclusions which contained very electron-dense spicular material and which were localized to muscle cells at the peripheries of the infarcts. These findings indicate that the spicular inclusions represent early stages in the process of mitochondrial calcification in myocardial infarcts. In contrast, calcium or other trace elements were not detected in moderately electron-dense amorphous inclusions which were present in mitochondria of muscle cells throughout the infarcts. With the tissue preparative techniques employed, the possibility cannot be excluded that the amorphous inclusions contained calcium, either in small amounts or in a readily diffusable state, in vivo. The data, however, are in accord with the previously advanced hypothesis that the amorphous inclusions represent precipitates of denatured mitochondrial protein formed during the evolution of irreversible cellular injury. This study provides further evidence that analytical electron microscopy can yield important information regarding the nature of various inclusions occurring in normal and diseased tissues.


Ultrastructural localization of viral antigens using the unlabeled antibody-enzyme method. Employing the unlabeled antibody enzyme method at the ultrastructural level, a comparison was made between preembedding staining and postembedding staining for the detection of viral antigens. The bacteriophage P1 absorbed to the surface of Shigella dysenteriae was used as a model system. Preembedding staining resulted in the specific deposition of peroxidase-antiperoxidase (PAP) complexes as an electron-dense coating around the viral heads. Disadvantages of the preembedding staining method included the agglutination of cells by the primary antiserum which produced a gradient of specific staining and the "bleeding" or migration of electron dense reaction product away from the sites of attached PAP complexes. The postembedding staining method had distinct advantages over the preembedding staining in that PAP complexes were deposited directly over exposed viral heads within the thin section. In addition, the specific immunostaining of viruses was uniform through the section and no artifactual migration of reaction product was observed.


Seroepidemiological study of HBV infections in Athens, Greece. A seroepidemiological survey of a sample-roughly representative by age and sex - of 2744 persons of the Greater Athens area revealed that hepatitis B virus (HBV) infections are highly endemic in Greece. Hepatitis B surface antigen (HBsAg) was detected in 72(2-6%) of them. The subtype was identified in 70 of the 72 carriers, and 69 were ay; the other was ad. Determinant w was present in all 61 that were capable of being typed further, and a23y(w) was twice as common as a11y(w). Antibody to HBsAg (anti-HBs) was found in 514 (18-7%) persons. The prevalence of the HBsAg rose rapidly with age, reaching peak values (5-3%) at 20-29 years, while anti-HBs reached its highest value (42-1%) in older age groups (50-59 years). The frequency of HBsAg was significantly higher in males (3-4%) than in females (1-5%). The percentage of infected persons who become chronic HBsAg carriers (12-3%) was found higher than in other developed populations. It was also found higher in children (25%) than in adults (5-5%) and in males (14-6%) than in females (8-1%). These data indicate that HBV infection in early life is a major risk factor in the development of HBsAg carriers and support the hypothesis that males are more likely to become HBsAg carriers than females.


Complement - fixation in the interaction of normal rabbit gamma G' globulin and its Fab' fragment. Complement consumption induced by Fab' fragments of rabbit IgG, irrespectively of their state of aggregation, requires the presence of homologous rabbit IgG. Up to a certain concentration of Fab' fragment, the amount of complement fixed is a linear function of the Fab' fragment dose, but further raising of the Fab'-fragment concentration does not result in complete complement consumption in the sample. The interaction between Fab' fragment and IgG is not strictly species specific.


Pulmonary changes in neonatal sepsis to group B beta-hemolytic Streptococcus: relation of hyaline membrane disease. The radiographic changes of hyaline membrane disease were seen in seven of eight infants with group B beta-hemolytic streptococcal sepsis. Hyaline membranes were found throughout the lungs of all six infants who were examined after death. The most striking finding was the presence of group B streptococci within the membranes of five infants. Streptococci were so numerous in one instance that they comprised the bulk of the membrane. Another infant uas thought to have classical hyaline membrane disease until numerous group B streptococci were found within the membranes when lung sections were examined with special stains. No other organisms were identified within the membranes of any infant. These findings suggest that infection with group B streptococcus may cause a syndrome clinically and radiologically indistinguishable from hyaline membrane disease.