Chemotherapy for testicular cancer: current status of the National Cancer Institute Combined Modality Trial. Twenty-five previously untreated patients with advanced testicular carcinoma were treated with a five-drug combination chemotherapy program containing cis-dichlorodiammineplatinum(II). This drug regimen was used as part of a trial testing the efficacy of cytoreductive surgery (15 patients) and in ten additional patients not eligible for the combined modality study. Eleven of 25 (44%) patients had a complete response and 11 of 25 (44%) patients had a partial response. Nine of 11 (82%) patients with minimal tumor burden upon initiation of chemotherapy had a complete response. With a median followup of 12+ months, none have relapsed. Complete remissions in patients with very advanced bulky disease were rare (two of 14 [14%] patients) and of short duration, with both patients relapsing. In addition to the usual hematologic toxicity, 14 of 25 (56%) patients developed hypomagnesemia and five of 25 (20%) patients developed systemic reactions to cis-dichlorodiammineplatinum(II) which prevented further drug administration.


Distribution and fine structure of the lymphatic system in the human testis. The distribution of lymph vessels in the human testis was investigated using ink injection methods, and light and electron microscopy. Lymph capillaries occur in the septula testis but are absent in the intertubular tissue. They consist of endothelial cells provided with an incomplete basal lamina and anchoring filaments of the adjacent connective tissue. Frequently, the endothelial cells are separated by gaps measuring up to 2 micron. The lymph capillaries of the septula testis are connected to lymph vessels in the rete testis and tunica albuginea. These vessels have occasional smooth muscle cells and valves. At the posterior margin of the testis, the network of lymph vessels merges into collecting ducts, which together with vessels derived from the rete testis are drained by the lymphatic system in the spermatic cord.


Localisation of nucleolus-organizing regions in interphase cells. A technique based on the use of silver solutions, which selectively stains the nucleolus-organising regions (NORs) in chromosomes, was applied to interphase Ehrlich tumour cells. The results indicate that nucleolar fibrillar centres correspond to the NORs.


ZIO staining in synaptic vesicles of the rat pineal nerves after inhibition of serotonin and noradrenaline synthesizing enzymes. Two compartments have been defined in monoaminergic synaptic vesicles: the core or central compartment, storage site for monoamines, and the matrix or outer compartment, of unknown function. The outer compartment reacts with the mixture of zinc iodide-osmium tetroxide (ZIO). This reaction is temperature and time dependent and may be abolished by -SH reagents. The effect of drugs inhibiting the synthesis of serotonin and noradrenaline (stored in the core) on the ZIO reaction in the matrix was studied in synaptic vesicles of rat pineal nerves. The inhibitors of monoamine synthesis abolish or decrease the ZIO reaction directly or in combination with the administration of tyramine. This effect is temperature dependent suggesting that the drugs act on different components of the matrix that react with ZIO at different temperatures. A comparison of the present results with those obtained with -SH reagents seems to indicate that the drugs assayed act, at least in part, by changing the accessibility of -SH groups in vesicle proteins. (An abstract of this paper was presented at the 7th International Congress of Pharmacology, Paris, 1978.)


Differential staining and segregation of parental chromosomes in mouse-rabbit hybridomas. Hoechst 33258 fluorescent staining can be coupled with G-banding to identify the chromosomal contribution of each parent in mouse-rabbit hybridomas. A fast and essentially complete segregation of rabbit chromosomes is obtained in these cells. The rabbit X chromosome is preferentially maintained in media imposing HGPRT activity for cell growth. Mouse-rabbit hybridomas, some of which secrete rabbit immunoglobulin chains, should be a convenient material for the identification of chromosomes governing rabbit Ig chain synthesis.


A cause of paired ventricular extrasystoles. Eight patients with ventricular extrasystoles are reported in whom coupling intervals of the extrasystoles to the proceding sinus beats were variable and in whom paired ventricular extrasystoles were occasionally seen. In all patients, paired ventricular extrasystoles were initiated only by comparatively late coupled ventricular extrasystoles. However, the interval between the first and the second of these paired extrasystoles was always much shorter than the coupling interval of this first extrasystole to the preceding sinus beat, so that the latter extrasystole often interrupted the T wave of the first one, indicating the R-on-T phenomenon. In two patients there was a gap between the ranges of coupling intervals for single extrasystoles and for the first ones of paired extrasystoles. These observations suggest the presence of longitudinal dissociation in the reentrant pathway as one of the causes of paired ventricular extrasystoles.


Evaluation of four "kit" immunoassay methods for determination of alpha-fetoprotein in serum during pregnancy. We evaluated four immunoassay methods for determination of alpha-fetoprotein in serum. All are commercially available in kit form in North American and we find them acceptable with respect to sensitivity, stability, precision, linearity, and analytical recovery. For the Dainabott method, the median values from 16 through 20 weeks of gestation are 34.4, 38.1, 42.1, 51.4, and 66.3 micrograms/L for each week, respectively. Corresponding values for the Amersham method are 27.9, 31.4, 34.9, 38.2, and 54.0 micrograms/L, respectively. For the Behring-RIA method they are 33.4, 37.4, 39.4, 49.9, and 66.3 micrograms/L, respectively. Results for four cases of confirmed neural tube defects are presented.


Kinetic aspects of the antigen-antibody reaction in various radioimmunoassays: effect of delayed addition of labeled or unlabeled antigens on sensitivity of assay. The kinetics of the antigen-antibody reaction were examined systematically in four kinds of double-antibody radioimmunoassay (RIA). In all the RIAs, the dose-response curves obtained on delayed addition by 24 to 48 h of labeled antigens (curves B), were shifted downwards and to the left of those obtained on simultaneous addition of the reagents (curves A), resulting in improved sensitivity of the assay. On the contrary, the dose-response curves obtained on delayed addition of unlabeled antigens (curves C), were shifted upwards and to the right of curves A, resulting in reduced sensitivity. In human thyrotropin (hTSH) RIA, curves B and C approached curves A very little, even after 168 h of incubation. A similar phenomenon was observed with anti-hTSH antisera from five different sources at two incubation temperatures, and the dilution curves of 125I-labeled hTSH and unlabeled hTSH appeared to be parallel. Therefore, the phenomenon observed with hTSH RIA could not be attributed to the assay conditions or to peculiar properties of the reagents used. In insulin RIA, the reversibilities of the shifts of curves B and C were slight but comparable to those observed in hTSH RIA. In 1-3,5,3'-triiodothyronine RIA, curves B and C gradually approached curves A on prolonged incubation and curves B became nearly identical with curves A after 98 h of incubation. On the other hand, in alpha-fetoprotein (AFP) RIA, curves B and C did not approach curves A, even on prolonged incubation for up to 288 h. The "equilibrium affinity constants" of the antibodies were of the same order of magnitude, thus it is unlikely that differences in the constants can account for the differences in the reversibility of these RIAs. In APF RIA, a significant amount of the antigen-antibody complex was precipitated without second antibody after centrifugation at 3000 X g. These findings suggest that the extent of dissociation of the immune complexes depends on their size, which in turn is related to the molecular weight of the antigen.


Decreased sialic acid and altered binding to lectins of purified alpha 2-macroglobulin from patients with cystic fibrosis. Purified preparations of plasma alpha 2-macroglobulin from patients with cystic fibrosis are shown to have normal amounts of total hexose but as much as 40% decrease in their sialic acid content. The binding of these preparations to concanavalin A and wheat-germ agglutinin was markedly reduced as compared to normal values in controls. Intermediate values were found in obligate heterozygotes. These results suggest a possible alteration in the carbohydrate moiety of alpha 2-macroglobulin in cystic fibrosis, presumably due to a defective posttranslational process.


Study of serum beta-2 microglobulin levels in breast cancer patients. Serum beta-2 microglobulin levels were measured in normal individuals and in breast cancer patients. It was observed that there was a significant rise in levels especially in advanced stages of the disease. It was concluded that patients with a beta-2 microglobulin/creatinine ratio of greater than 3.8 were likely to have metastatic breast cancer.


Selective production of human antigen specific helper factor from normal volunteers: implications for human Ir genes. Based on previous systems for generating helper cells and factors from mouse spleen cell cultures, an in vitro system for the production and detection of human helper factors to the synthetic polypeptide antigens (T,G)-A--L and GAT10 was developed. The factors are made by human peripheral blood leucocytes and are antigen-specific, as judged both by functional criteria and specific binding and elution from antigen columns. Out of the first six volunteers studied two were high responders to (T,G)-A--L, but non-responders to GAT, two responders to GAT but not (T,G)-A--L. One subject made factors to both antigens and the sixth reacted to neither. The antigens chosen are known to be under MHC-linked immune response (Ir) gene control in all animal species tested, with some strains being responders while others are not. The selective responsiveness, different between individuals, thus suggests that the response to these antigens in man is under Ir gene control. Because of the small size of the sample initially studied and HLA typed, it was not surprising that there was no clear-cut association of response with any particular histocompatibility type at the HLA-A, B, C or D locus.


Antigens of the human trophoblast plasma membrane. Using purified plasma membranes of human syncytial trophoblast as immunogen, we have raised heterologous antisera which reacted by indirect immunohistological methods with the surface of the trophoblast. This antigenic activity appears to be absent from other adult and foetal tissues. Its absence from maternal sera and its resistance to mild detergents treatment suggest that it is intimately associated with the surface membrane of the syncytial trophoblast. This antigen was found to be immunologically different to a number of proteins, including those known to be produced by the trophoblast. The identification of this antigen may indicate a potential line of research for the immunological regulation of human fertility.


Antigen-specific detection of HBsAG-containing immune complexes in the course of hepatitis B virus infection. In recent studies extrahepatic manifestations of viral hepatitis have been recognized as immune complex diseases. Hepatitis B surface antigen (HBsAg) has been successfully identified in immune complexes, but the pathogenic role of HBsAg-containing immune complexes (IC) remains questionable. The subject of the present study was the antigen-specific determination of IC in the course of hepatitis B virus infection using a new HBsAg-specific IC test (Pernice & Sedlacek, 1978). This test is based on the following principle: rabbit anti-HBs-coated polystyrole test tubes are incubated with the IC-containing test sample. The HBsAg-containing IC bind to the solid phase by their free antigenic determinants. There they can be quantified using a peroxidase-labelled anti-human IgG antibody. A good correlation was found between the level of HBsAg-containing immune complexes and the clinical state of six patients in a follow-up study. IC could be detected simultaneously with HBsAg and either decreased or disappeared before the occurrence of free anti-HBs. In the sera of an additional twenty-eight patient suffering from chronic active hepatitis, HBsAg-containing immune complexes were detected in 85% of cases. One patient suffering from polyarteritis nodosa was also positive. Occasionally, extremely high levels of IC were found in the course of these diseases.


Chemotherapy of Hodgkin's disease.

Chemotherapy of Hodgkin's disease. During the past 10 to 15 years, there have been dramatic improvements in the prognosis of patients with advanced Hodgkin's disease resulting from the development of combination chemotherapy. The long-term follow-up of patients with advanced disease treated with MOPP indicates that 66 per cent of those achieving complete remissions have not relapsed and remain off therapy at five and 10 years follow-up. Attempts to make additions, subtractions or modifications in the MOPP regimen have not generally improved the overall survival rate or reduced the toxicity. Development of non-cross-resistant combinations provides a rational approach to the treatment of primary induction failures and those who relapse after short initial remissions. Alternating sequence induction chemotherapy regimens show considerable promise. The use of combined modality approaches to the treatment of localized IB--IIIB Hodgkin's disease has made a significant impact on disease-free survival but has yet to alter actual survival. Combined modality approaches in advanced disease are encouraging but as yet long-term follow-up is not available to assess their worth vis-à-vis existing therapies for advanced disease. Improved survival has brought about concern over late complications, particularly second malignancies. These complications require careful monitoring and must be balanced against any survival benefit seen from combined modality approaches. However, the fact that patients now survive long enough to be potentially at risk for such long-term complications is a testament to the efficacy of modern day aggressive therapy.


N-Acetylprocainamide pharmacokinetics in functionally anephric patients before and after perturbation by hemodialysis. NAPA pharmacokinetics were studied in 6 functionally anephric patients. Distribution and nonrenal elimination of this drug were found to be the same as in individuals with normal renal function but renal clearance was reduced, resulting in a mean elimination t 1/2 of 41.9 hr (6.2 hr in normal subjects). Renal clearance of NAPA correlated well with ClCr. Dialysis removed NAPA from both red blood cells and plasma and increased ClT approximately fourfold. Dialysis itself resulted in a 77% reduction in ClS that limited the total amount of NAPA removed by this procedure. This reduction in ClS was sustained for at least 3 hr after dialysis and attenuated rebound in plasma NAPA concentrations.


Cardiac rhythm in tricyclic antidepressant poisoning. One hundred adult patients with tricyclic antidepressant poisoning were studied by clinical observation and EKG monitoring. Sinus tachycardia was common but dysrhythmias were uncommon and unimportant. EKG monitoring did not influence the treatment of these patients and the value of its routine use in tricyclic antidepressant poisoning is questioned.


Transport of vitamin D metabolites.

Transport of vitamin D metabolites. Vitamin D and its metabolites are bound to an alpha globulin (DBP) in human serum. This carrier protein binds 25-OHD and 24,25(OH)2D with higher affinity than vitamin D or 1,25(OH)2D, but the binding is highly specific for the vitamin D structure. The carrier mechanism appears to be unique in that it is a high affinity and high capacity system, capable of binding as much as 120,000 IU of biological activity per liter of plasma. DBP is apparently identical to group-specific component, and a DBP-deficient state has not been identified among approximately 75,000 human sera examined thus far. The liver appears to be the site for synthesis of DBP, and serum DBP levels are increased during pregnancy and during estrogen-progesterone therapy. However, in a variety of disorders of mineral homeostasis, serum DBP levels are normal. Two tissue binding proteins for vitamin D metabolites have been identified. One protein, sedimenting at 5-6S, has been found in all nucleated tissues, and exhibits a ligand preference similar to DBP. This tissue binding protein appears to be a complex of serum DBP with a tissue protein which is heat-labile, and which shows no capacity for binding vitamin D sterols. The physiologic role, if any, for this complex of serum DBP and tissue protein is not presently understood. The other tissue binding component is a 3-4S protein found only in recognized target tissues, and which shows a high binding affinity and specificity for 1,25(OH)2 D. It appears to be the receptor in the receptor-1,25(OH)2D migration to the nucleus, leading to the biosynthesis of mRNA which codes for products which affect calcium and phosphorus transport in target tissues.


Buridan's ass: one man's view of the immune system. This essay offers a highly biased and prejudiced interpretation of the immune response. The major points of this interpretation are: B and T lymphocyte diversity is generated by rearrangements within and among existing receptorcoding genes; the T cell receptor is a molecule genetically more closely related to immunoglobulins than to the products of the major histocompatibility complex (MHC); the course of B and T lymphocyte differentiation is largely the same except for a final processing of T lymphocytes in the thymus; the processing is directed by MHC molecules of the thymus epithelium; Ir and Ia genes are identical; and the function of MHC polymorphism is to generate diversity at the population level.


[Alpha-1-fetoprotein in tumour tissue (author's transl)]. Using the indirect immunoperoxidase method, alpha 1-fetoprotein (AFP) can be demonstrated in paraffin sections of tumours by light-microscopy. Among 160 tumour preparations it was demonstrated in five of six endodermal sinus tumors, 19 of 40 teratomas, the three hepatoblastomas and 9 of 16 primary liver-cell carcinomas. Among 15 rhabdomyosarcomas, 15 Wilms tumours, 15 neuroblastomas and 15 other malignant tumours only one rhabdomyosarcoma contained AFP. No AFP was demonstrated in 35 various benign tumours and various other human tissue samples. Distribution of AFP-producing cells in tumour tissue is generally focal samples. In malignant teratomas AFP was found only in areas which corresponded to an endodermal sinus tumour or an embryonic carcinoma. The practical significance of the results consists of (1) demonstration and localisation of AFP in tumour tissue; (2) aiding the differential diagnosis and classification of tumours and their metastases; and (3) recognition of pathogenetic connection between embryonic carcinoma and endodermal sinus tumour.


Foveal versus peripheral retinal responses: a new analysis for early diagnosis of multiple sclerosis. Visual potentials evoked by brief flashes (VEPs) were recorded in 85 patients with MS, 30 healthy controls and 25 neurological patients without demyelinating diseases. In a group of 44 patients, diffuse field stimulation was used, resutling in 15 altered responses (34%). In a second group of 41 patients with superimposable characteristics including age, sex, diagnosis of MS (definite, probable and possible) and score on the Rose's scale, the separate responses of central and peripheral retina were analysed. A total of 30 abnormal VEPs were recognized (73%) with the following distribution: 16 foveal VEPs (78%), 4 peripheral responses (13%) and 26 centreperiphery latency differences (CPLD, 60%). Twenty subjects of Group A with normal or slightly altered full field VEPs were retested with both stimulating methods with the following result: 3 altered full  field VEPs (15%) and 15 abnormal CPLD (75%). With diffuse field stimulation, the correlation between the VEP scores and the results of other examinations (visual, pyramidal, etc.) were insignficant; on the contrary, the CPLD was clearly correlated with the involvement of the visual and other systems in the fluctuating clinical course of MS.


Short latency mechanically evoked somatosensory potentials in humans. Somatosensory potentials evoked by mechanical stimulation were recorded by surface electrodes over (1) the digital nerves in the index finger, (2) the median nerve at the wrist, (3) the median nerve near the axilla, (4) the brachial plexus, (5) the cervical cord at CII, (6) the scalp overlying the somatosensory cortex. Nerve conduction velocities varied inversely with age and ranged from 43 to 68 m/sec. Mechanically evoked potentials recorded from the electrodes overlying the digital nerves were an artifact of the finger movement. All other electrode configurations recorded potentials comparable to those evoked by electrical stimulation of nerves. These mechanically evoked potentials could prove useful in the assessment of clinical disorders of somatosensory function from receptor to cortex in man.


Dipole-like neuronal sources of theta rhythm in dorsal hippocampus, dentate gyrus and cingulate cortex of the urethane-anesthetized rat. Spatial distribution of theta activity was investigated in the dorsal hippocampal formation and overlying neocortex of the urethane-anesthetized rat. Laminar phase profiles from semi-microelectrode penetartions showed approximately 180 degrees phase shifts combined with small amplitude values in stratum radiatum of CA1, instratum moleculare of the dentate gyrus and in layer V/VI of the cingulate cortex at theta peak frequency. Evidence has been presented that layers of neurons in CA1, in the dorsal granular layer and in the cingulate cortex are the sources of dipole-like theta field potentials. A strong linear relationship between the neuronal theta sources in hippocampal CA1, dentate area and cingulate cortex was found.


L-tryptophan and sleep in healthy man.

L-tryptophan and sleep in healthy man. The effect of L-tryptophan on night-time and day-time sleep (from 14.00 h) sleep was studied in six healthy males aged between 20 and 30 years. The doses used in the night-time studies were 2, 4 and 6 g, and in the day-time studies 1, 2 and 4 g. It was not possible to establish an effect of L-tryptophan compared with placebo on night-time sleep, but analysis of the sleep measures with 4 g compared with placebo and the other doses of L-tryptophan considered together suggested reduced awakenings, increased stage 3 and an increased percentage of REM sleep. With 4 g L-tryptophan there was an increase in the duration of stage 3 of day-time sleep compared with placebo. The studies provide marginal evidence that REM sleep may be modified by L-tryptophan in man, though the evidence is somewhat stronger that SWS may be increased. The effect on REM sleep may involve circadian mechanisms. The hypnotic activity of L-tryptophan per se is limited and uncertain.


Conditioning auditory stimuli and the cutaneous eyeblink reflex in humans: differential effects according to oligosynaptic or polysynaptic central pathways. Electromyographic activity of orbicularis oculi muscles in humans was elicited by percutaneous electrical stimulation of the supraorbital brahcn of the trigeminal nerve. The reflex consists of an early brief ipsilateral R1 and a later prolonged consensual R2. The threshold for R1 was considerably elevated compared to that of R2. In one experiment brief acoustic stimuli, at 70 dB SPL, were presented at various intervals, from 5 to 800 msec, prior to the eliciting stimulus. In a second experiment similar stimuli, with intensities varying from 30 to 70 dB SPL, were given at the fixed lead time of 100 msec. In each experiment the preliminary acoustic stimulus enhanced R1 and depressed R2. Potentiation of R1 developed more rapidly than did depression of R2 and exhibited an early and a late peak, whereas depression had a single intermediate trough. Both effects linearly increased with increases in the intensity of the acoustic prepulse. These restuls are discussed in relation to the neuronal circuits responsible for the expression of the two reflex components.


Changes in human event related potentials with prolonged delta-9-tetrahydrocannabinol (THC) use. Auditory event related potentials and contingent negative variation (CNV) were recorded in a series of behavioral tasks from 27 hospitalized human volunteers before, early during (1--3 days), late during (10--14 days) and after chronic THC intoxication at two dose levels (70--90 mg/day or 180--210 mg/day in divided doses). Twelve additional hospitalized volunteers served as a control group. The N1 component of the auditory event related potential in all tasks was depressed when tested early during 180--210 mg/day THC administration. No N1 changes were seen for the subjects receiving 70--90 mg/day or the control subjects. Ten to 14 days during the chronic THC perioe CNV was depressed in all tasks except the control task for both doses at the early THC testing. Only at the most difficult task at the higher of the doses was the CNV depressed at the test late in the THC period. The changes seen in the N1 and the CNV may be indicative of different aspects of a common attentional alteration during stimulus processing. The results on our more difficult tasks suggest the alteration may continue in complex stimulus processing tasks in heavy marijuana users.


Period-amplitude analysis of EMG from slow and fast extensors of cat during locomotion and jumping. A real-time period-amplitude analysis program for EMG involving the detection of events consisting of the period and amplitude difference between maxima and minima was used to study the activity patterns of fast and slow, hind and forelimb extensors during treadmill locomotion and jumping. Events were stored in a two-dimensional table providing an easily interpreted quantification of frequency and amplitude parameters of single EMG bursts which were characterized over a range of movements by the mean frequency and amplitude. Periods between zero crossings of the original signal were also stored. Both mean frequency and mean amplitude remained relatively steady for the slow extensors throughout the range of movements, while the fast extensors exhibited an increase in both mean frequency and mean amplitude during the same movements. The results show that fast extensors of the hind and forelimb follow the kinetic demands of the movement, while the activity pattern of the slow extensors is constant and independent of the movement kinetics. Slight discontinuities in these trends are discussed in their possible relation to the dynamics of msucle recruitment. Period-amplitude analysis provides a useful method of quantifying the raw EMG while retaining much of the original information of the signal.


Medial thalamic permanent electrodes for pain control in man: an electrophysiological and clinical study. In 7 patients with medial thalamic electrodes implanted for relief of chronic pain, clinical observations and electrophysiological recordings showed: (1) Clinically, paresthesia in the pain area and contralateral half of the body was reported as well as some unpleasant side effects. The EEG was not changed during medial thalamic stimulation. (2) Threshold for non-painful and painful perception of electrical stimuli were not significantly raised by contralateral medial thalamic stimulation. (3) After electrical median nerve stimulation, evoked thalamic potentials started after 17 msec. With ipsilateral stimulation the early components (up to 40 msec) showed longer latencies and the late components slightly shorter latencies as compared to the contralateral stimulation. (4) Median nerve evoked cortical SSEPs are not significantly changed either by a 500 msec lasting conditioning stimulation of the dorsal columns or by such stimulation of the medial thalamic structures. A facilitation at about 90 msec was found in the cortical SSEP after medial thalamic conditioning in only one of three patients tested. (5) Visual and auditory evoked potentials can be recorded in the medial thalamus as well.


Spontaneous interictal spiking in the awake kindled rat. The spontaneous interictal spike (SIS) noted in EEG recordings is a signal of the abnormally excitable behavior of neurons in an epileptic focus. This study provides a detailed, quantitative, temporal and spatial profile of SIS in the amygdaloid kindled rat. The reproducible nature of the development and decline of SIS in kindling provides a useful model for biochemical study of the mechanisms that initiate and regulate SIS.


Wakefulness-sleep modulation of thalamic multiple unit activity and EEG in man. Quantitative changes in the number of EEG waves and MUA neuronal spikes of various thalamic nuclear groups were studied during steady and transitional wakefulness-sleep states in a group of patients with implanted electrodes, used as an electrophysiological procedure for the surgical treatment of unilateral tremor and rigidity. Thalamic nuclei included ventroposterolateral (VPL), ventrolateral (VL) and centromedian (CM). Steady states included initial wakefulness (W1), slow wave sleep (SWS) I, II, IV and IIL, paradoxical sleep phasic (PSP) and tonic (PST) and final wakefulness (W2). Transitional states included transition from SWS to PSP (transition) and from sleep to W2 (arousal). (1) There was a significant correlation in EEG and MUA between different thalamic nuclei simultaneously recorded in the same patient during different steady-states. (2) Thalamic MUA significantly decreased from W to SWS, increased from SWS to PS and SWS to W2, while thalamic EEG significantly decreased from W to SWS, increased from SWS to W2 but was not significantly modified from SWS to PS, consecutive steady-state shifts. (3) Thalamic MUA significantly decreased while EEG remained unchanged from PSP to PST consecutive steady-state shifts. (4) Both thalamic MUA and EEG significantly decreased from SWS II to IIL while they were not systematically modified from W1 to W2 indirect steady-state shifts. (5) Thalamic MUA significantly increased 1 sec before transition and arousal while EEG increased 5 sec after arousal and did not change during transition.


Brain stem auditory evoked potentials and blink reflexes in quiescent multiple sclerosis. Brain stem auditory evoked potentials (BAEP) and blink reflexes (BR) were studied in 25 patients with multiple sclerosis (MS), the diagnosis being definite according to McAlpine's criteria, in the quiescent phase, without signs of brain stem involvement. BAEP abnormalities were found in 64% of the cases and BR abnormalities in 60%. A good correlation between the two tests was found in most patients. The abnormalities consisted of delayed latencies and/or high intraindividual variability in shape and latency of BAEP and BR components. It seems that demyelination of brain stem pathways results not only in conduction slowing but also in more serious dysfunction of the generators of the evoked components.


True checkerboard pattern reversal with light-emitting diodes. The use of rectangular red light-emitting diodes permits construction of a true checkerboard pattern-reversal stimulator, which given evoked responses comparable in amplitude and wave form to those obtainable by a TV pattern generator. The device interfaces easily with commercial avergers and allows the incorporation of several features that may increase the sensitivity of visual evoked responses in screening for multiple sclerosis.


Serial quantitative electroencephalography. Repeated EEG examinations are often used in diagnostically difficult cases to assess whether an EEG abnormality is progressive, regressive, or stationary. The conventional visual method is, however, poorly adapted to consider the variability when consecutive EEG recordings are compared. Using computer analysis of EEGs, the feature of the recordings as well as their variances can be measured, and the differences between the recordings can be tested statistically. The aim of the present study was to develop a practicable method for quantitative serial EEG examinations. Twenty EEGs from 5 volunteers were recorded to obtain reference data. The data acquisition was automated to obtain data without visual bias. The previously developed method using age ratios was employed for the primary analysis. The changes of the EEG activity during the observation period were evaluated statistically. Differences between consecutive EEGs could be observed even in normal subjects but the number of 'false positive' findings was reduced by the automatic data selection, by the choice of proper measures, and by adequate statistical treatment. The diagnostic value of the method was tested in 59 EEGs from 13 patients with well-defined brain disorders. Most EEG changes which could be attributed to the development of the disease were discovered and labelled as statistically significant.


A portable pulse-interval modulation telemetry/multiplexing EEG recording system for use in the home. A portable 6-channel recording system employing time-division multiplexing with cable or radio-telemetry has been developed. The system is very compact and has proved to be reliable when used in a domestic environment. The use of multiplexing, in general, permits an expansion in the number of available channels and a reduction in equipment and tape costs, while the use of pulse interval modulation, in particular, overcomes tape recording limitations which may be encountered with the more common amplitude modulation techniques.


A 16-fold semi-microelectrode for intracortical recording of field potentials. A 16-fold semi-microelectrode with 10 x 10 sq. microns contacts at distances of 150 microns, produced by means of thin-film technology, is described. These small dimensions are required to study the laminar electrophysiological differences within the cerebral cortex.


Event-related potential changes in chronic alcoholics. Ten chronic alcoholics (10+ year alcoholic drinking history) and ten age and sex matched controls were tested on an ERP paradigm which elicited a large P3 component. The N1 and P2 sensory evoked potential components did not differ in amplitude or latency between the two groups. The latency of the P3 was significantly longer for the alcoholics than the controls for both a response and non-response stimulus. This finding is consistent with the results seen in a variety of dementias and is offered as evidence of the dementing effects of prolonged alcoholism in this group of subjects. While the P3 latencies were prolonged for the alcoholics, their reaction times were not different from the controls. Single trial analysis using Woody's adaptive filter also demonstrated that the single trial estimates for the 3 latency were significantly prolonged for the alcoholics. The single trial correlation between the P3 latency and each trial's corresponding reaction time was significantly greater for the alcoholics than for the controls.


Hyperbaric hyperreflexia: tendon-jerk and Hoffmann reflexes in man at 43 bars. Tendon jerk (TJR) and Hoffmann (H) reflexes of the soleus muscle were studied in two men during a 26-day simulated oxygenhelium dive to a maximum pressure of 43 bars. The amplitude of the TJR response was observed to increase markedly after compression and also at the end of decompression. This biphasic pattern of enhanced reflectivity was reproduced by synchronised but much smaller variations in H reflex response. The positive facilitatory effects of applying Jenkdrassik's manoeuvre were reduced and the negative effects increased during hyperbaric exposure. Excitability cycles (twin-pulses methods) revealed modifications in one subject similar to those observed in clinical hyperreflexia, namely a shortening of Phase III and an enhancement of Phase IV. The other subject exhibited a notable depression of Phase IV. Abnormal EMG recordings are described including randomly-triggered slow wave potentials with no mechanical effects, and fixed, long latency late responses of a reflex nature with a definite mechanical effect. The contention that alterations in vestibulo-spinal relations may result in the release from tonic presynaptic inhibitory control of myotatic reflex arc excitability is discussed as a partial explanation for these and related findings.


EEG patterns during 'cognitive' tasks. I. Methodology and analysis of complex behaviors. This paper presents a methodology which uses nonlinear pattern recognition to study the spatial distribution of EEG patterns accompanying higher cortical functions. The multivariate decision rules reveal the essential EEG patterns which differentiate performance of two tasks. Cross-validation classification accuracy measures the generality of the findings. Using this method, EEG patterns were derived from a group of 23 adults during performance of several complex tasks, including Koh's block design, writing sentences, mental paper folding, and reading silently. These patterns discriminate between the tasks, are consistent with, and extend the results of, visual EEG interpretations and univariate analysis of spectral intensities. Since writing sentences could not be distinguished from mere scribbling, it is unclear whether the EEG patterns found to distinguish complex behaviors were related to the cognitive components of tasks, or to sensory-motor and performance-related factors.


EEG patterns during 'cognitive' tasks. II. Analysis of controlled tasks. This experiment was designed to distinguish possible EEG correlates of the cognitive components of tasks from EEG patterns associated with stimulus characteristics, limb and eye movements, and performance-related factors such as subjects' ability and effort. Thirty-two right-handed adults each performed 30 trials, lasting 6-15 sec each, of four simplified, controlled tasks: mental rotation of geometric forms, serial addition of a column of signed digits, substitution of letters with subsequent word recognition and visual fixation. The first three tasks could not be differentiated from each other. Each of these tasks could be differentiated from visual fixation by approximately 10% generalized reductions in alpha and beta band intensities, and slight increases in theta band intensities frontally and occipitally. We conclude that the EEG patterns which differentiated the complex tasks described in Part I were due to inter-task differences in stimulus characteristics, efferent activities and/or performance-related factors, rather than to cognitive differences. With these controls, no evidence for lateralization of different types of cognitive activity was found in the EEG.


Effects of stimulus frequency on the blink reflex during rest and a task. Blink reflexes were studied in 84 healthy subjects, applying 4 different stimulation frequencies. The experiments were subdivided into 3 periods: rest, binary choice task and final rest period. Independent of the applied stimulation frequency, an increment of R1 during the task was found, and it was concluded that R1 amplitude reflects the state of arousal of the subject. Within each of the 3 periods, R1 amplitudes decreased independently of the applied frequency, while R2 amplitudes decreased dependent on the applied frequency, i.e. at lower frequencies a smaller decrement in time could be observed. It was concluded that R2 habituated at frequencies lower than those so far reported.


Dependence of refractory period measurements on conduction distance: a computer simulation analysis. Computer simulations were used to study the recovery of conduction velocity in the temporal and spatial domains during the relative refractory period. The impulse exhibits increased duration and reduced amplitude during the relative refractory period as well as a reduced conduction velocity. Changes in conduction velocity are reflected by a deviation from linearity in the plots of latency variation versus interstimulus interval and of interval-between-impulses versus intersimulus interval. The latency increase for the second impulse is monotonically related to interstimulus interval, and is maximal at the beginning of relative refractory period. The magnitude of the latency increase is also related monotonically, but not in a linear manner, to conduction distance. The distance-dependence of latency variation during the relative refractory period is most marked close to the site of impulse initiation. These findings may be relevant to relative refractory period measurements in clinical neurophysiology.


Procaine-induced seizures in epileptic monkeys with bilateral hippocampal foci. Intravenous procaine HCl given at low doses (0.5-2.5 mg/kg) to two monkeys with bilateral alumina hippocampal foci depressed interictal spiking or had little effect. At 5.0 mg/kg unilateral limbic activation occurred. At 10.0 mg/kg unilateral or bilateral limbic activation and generalized seizures could be evoked within 3-10 min. At higher doses (15 and 20 mg/kg) bilateral limbic activation or brief (one min) generalized seizures occurred. The unilateral-onset psychomotor seizures were not identical to spontaneous psychomotor seizures, and the generalized seizures never occurred spontaneously in these monkeys. However, these results do indicate that procaine challenges may selectively activate limbic epileptogenic areas without activation of debilitating generalized tonic-clonic seizures.


A pulsed integrator for EMG analysis.

A pulsed integrator for EMG analysis. A novel device is described which converts EMG signals to an output which is proportional to the area under the rectified curve in discrete time intervals. This is achieved by resetting a long time constant integrator at the desired rate and saving each integral in a sample/hold circuit for output during the subsequent integration period. This output is particularly useful for quantifying fast reflex events since the signal is integrated without leakage in each period and leaves no residual signal in subsequent periods.


Comparison of interwave latencies of brain stem auditory evoked responses in narcoleptics, primary insomniacs and normal controls. A study of brain stem auditory evoked responses (BAER) was carried out in 10 narcoleptics, 10 primary insomniacs and 10 normal controls to determine if a neurophysiologic abnormality could be detected in these primary sleep disorders. The mean interpeak conduction times of Wave I-III, III-V and Iv were compared between the following groups: normal controls awake and in monitored sleep; narcoleptics awake and in monitored sleep, normal controls awake and narcoleptics awake; normal controls awake and insomniacs awake; narcoleptics awake and insomniacs awake; narcoleptics with cataplexy (n = 6) awake and narcoleptics without cataplexy (n = 4) awake. No significant differences were found which suggests that these sleep disorders represent dysfunctions which do not involve brain stem structures subserving the BAER.


Convulsive effects of 3-aminopyridine on cortical neurones. The authors examined the effects of 3-aminopyridine (3-AP) on pyramidal tract neurones in the motor cortex of anesthetised cats. 3-AP proved to be a strong convulsive agent. Local application of 3-AP in concentrations of 15-20 mM resulted in typical paroxysmal alterations of neuronal activity: (a) enhancement of background firing, (b) augmentation of EPSPs, (c) depression of IPSPs and (d) single and serial paroxysmal depolarization shifts (PDSs). Serial PDSs were reflected by large synchronous waves in the surface ECoG.


A microdrive for use with glass or metal microelectrodes in recording from freely-moving rats. A new subminiature microdrive assembly is described for electrophysiological recording from behaving rats. This very small, lightweight system allows excellent precision in electrode placement and can maintain stable recordings over extended periods. Since the electrode is nonrotating, tissue damage is minimized. Either metal or glass microelectrodes may be used with the system, offering the possibility of iontophoresis for cell marking or neuropharmacological manipulations.


Properties of androphilic proteins in cytosols of human benign prostatic hypertrophy. Androphilic proteins in the cytosol from the human benign prostatic hypertrophy are separated into two fractions by Sephadex chromatography; void volume fraction and IgG fraction which was eluted near the site of hIgG. In the present study, properties of these two androphilic proteins were compared. Association constants of these proteins were in the order of 10(9) M-1. However, the binding capacity of the former was smaller than that of the latter. These two androphilic proteins well bound to nuclei, and the high-affinity and saturable binding to nuclei was observed in the 3H-dihydrotestosterone-IgG fraction complex, while binding of 3H-dihydrotestosterone-void volume fraction complex to nuclei was low affinity and unsaturable. The binding of the complexes to chromatin seems to be of low affinity and nonsaturable. These androphilic proteins did not bind to calf thymus DNA. Salt extractability of the bound void volume fraction after incubation with nuclei was not different from that of the bound IgG fraction. It was observed that the chromatographic behavior of the androphilic protein in IgG fraction was changed after incubation with nuclei.


The influence of chemotherapy on the distribution of 57Co-bleomycin in chemically induced squamous cell carcinoma of mouse skin. Organ distribution and tumor uptake of 57Co-bleomycin (BLM) were examined in NMRI mice without tumor and with chemically (DMBA-) induced squamous cell carcinoma. In these tumors high accumulation of 57Co-BLM was recorded shortly after injection of 57Co-BLM and low uptake 24--48 h after application. Two groups of tumor-bearing mice received unlabeled bleomycin as a cytostatic in two different doses. Uptake of 57Co-bleomycin was reduced. The study suggests that in patients currently or previously under chemotherapy, 57Co-bleomycin tumor scintigraphy may lead to false negative results.


Chemical and biological properties of bleomycin labelled with 57Co. The chemical composition of bleomycin labelled with 57Co and its distribution and kinetics in the organs of tumour bearing rats were studied. For separating bleomycin, the chromatography on a CM-Sephadex C-25 column with a gradient of ammonium formate was found to be most suitable. It is also possible to use paper chromatography in 10% NH4Cl and thin layer chromatography in 10% ammonium acetate with methyl alcohol 1:1. The fractions obtained by the chromatographic separation were identified as A2, B2 and A1. Labelling with 57Co given rise to a more complicated mixture of labelled compounds, all of them differing chromatographically from the original non-labelled components. Using carrier-free 57Co two main radioactive compounds are formed apparently corresponding to the fractions A2 and B2 containing one atom of Co per molecule. The amount of another two labelled components grows progressively with increasing amount of carrier CoCl2. Since their chromatographic properties are even more different from those of the original compounds, it was concluded that they probably contain two atoms of Co in their molecules. During distribution studies, rapid excretion of labelled bleomycin into the urine and stool was observed. Increased activities were retained in the liver, kidney, intestine and tumour.


Evaluation of cold areas on the thyroid scan with 99mTc-bleomycin. The value of 99mTc-bleomycin scintigraphy in the evaluation of cold areas on a 99mTc-pertechnetate scan was investigated in 35 patients. A histological diagnosis was obtained in 31 patients. Only one of the six malignant lesions was considered as positive and two as doubtful. Of the remaining 25 patients with non-toxic colloid goitre, one was clearly positive and four were doubtful. These results indicate that 99mTc-bleomycin scintigraphy is not a valuable procedure in the further evaluation of patients with cold lesions.


Correlations between age-dependent protein and lipid concentrations in plasma and platelet functions in children. The correlations between the levels of various plasma proteins and lipids and platelet function on glass and platelet factor 3 (PF 3)-availability in children of different age-groups were investigated. Several statistically significant positive and some significant negative correlations were found. Although conclusions based solely on such correlations should be considered with reservation, in our opinion the following factors should stimulate platelet function: prealbumin (adhesion and PF 3-availability in all age-groups, aggregation--specifically for children in puberty); alpha 1-antitrypsin (PF 3-availability); alpha 2-macroglobulin (platelet spreading capacity, PF 3-availability); plasminogen (platelet adhesion and aggregation--specifically for boys in puberty); caeruloplasmin (number of "free adhering platelets" spreading capacity); lysolecithin and lecithin (time-dependent increase of spontaneous platelet adhesion and aggregation, PF 3-availability); and free fatty acids (FFA) (PF 3-availability). Plasminogen and complement component C'3 show a negative relationship to the time-dependent increase of spontaneous platelet adhesiveness and aggregability in platelet-rich plasma.


The Langer-Giedion-syndrome (tricho-rhino-phalangeal syndrome, type II). A 15-year-old girl with the Langer-Giedion-syndrome (tricho-rhinophalangeal syndrome, type II) is presented. The features are multiple exostoses, sparse, fine hair and an abnormal face and hands. X-ray examination reveals exostoses and abnormal epiphyses. In this case intelligence is normal; there is severe sensorineural deafness (60--80 dB hearing loss). The genetic aspects of the syndrome are discussed.


Enhanced expression of HLA antigens and beta 2-microglobulin on interferon-treated human lymphoid cells. Human leukocyte interferon increased the expression of HLA-A, B antigens and beta 2-microglobulin on two lines of human lymphoblastoid cells and on peripheral blood lymphocytes. No effect was observed on the expression of HLA-DR antigens.


Occurrence of beta 2-microglobulin in mammalian lymphocytes and erythrocytes. Cell-associated beta 2-microglobulin (beta 2m) has been studied by quantitative techniques in lymphocytes and erythrocytes of guinea pig, rabbit, rat and man. beta 2m was solubilized by sonication followed by treatment with acid, deoxycholate or thiocyanate, and then determined by radioimmunoassay. The average number of beta 2m molecules per lymphocyte, estimated after acidification, varied between 0.89 X 10(5) and 7.1 X 10(5). Rat erythrocytes contained 3.0 X 10(3) molecules per cell, whereas no beta 2m was detected in red blood cells from the three other species. A relatively large part of beta 2m in the mammalian lymphocytes and in rat erythrocytes was present on the cell surface according to a radioimmunoassay procedure using anti-beta 2m antisera that had been absorbed with the blood cells analyzed.


Serological analysis of antigen-specific helper factors specific for poly-L(Tyr, Glu)-poly-DLAla--poly-LLys [(T, G)-A--L] and L Glu60-LAla30-LTyr10 (GAT). In vitro prepared antigen-specific helper factors reactive to the synthetic polypeptide antigens poly-L(Tyr, Glu)-poly-DLAla--poly-LLys [(T, G)-A--L] or LGlu60-LAla30-LTyr10 (GAT) and bearing Ia determinants were analyzed serologically to determine the nature of the Ia determinants they expressed. I subregion-specific mouse anti-Ia antisera were used, and showed that (T, G)-A--L-specific helper factor (HF) contains I-A subregion-controlled determinants, whereas GAT-specific HF carries I-J subregion-controlled antigens. This unexptected finding was confirmed in both the H-2k and H-2 b haplotypes, using a variety of anti-I-J antisera. Rabbit anti-Ia antisera also reacted with both HF which raised the possibility that the Ia determinants on HF may be carbohydrate in nature. The fact that HF has a low molecular weight and yet contains Ia determinants, antigen-binding capacity and idiotypic markers is compatible with this interpretation.


Characterization of a monoclonal anti-beta 2-microglobulin antibody and its use in the genetic and biochemical analysis of major histocompatibility antigens. A monoclonal anti-beta 2-microglobulin (BBM.1 antibody) was produced by cell fusion between the mouse myeloma, P3-X63-Ag8, and spleen cells from a BALB/c mouse immunized with Molt 4, a human T cell line. BBM.1 antibody was fully inhibited by soluble beta 2-microglobulin and purified HLA-A, B antigens and reacted with human-mouse somatic cell hybrids only if they had chromosome 15 and expressed human beta 2-microglobulin. It was cytotoxic in complement-dependent lysis and of the IgG class. BBM.1 and a monoclonal anti-HLA-A, B, C glycoprotein antibody, W6/32 (Barnstable, C. J. et al., Cell 1978. 14:9.), were used to quantitate relative amounts of beta 2-microglobulin and HLA-A, B, C glycoproteins on different human cell types. Thymocytes and the Molt 4 cell line showed a considerable excess of beta 2-microglobulin over HLA-A, B, C glycoproteins, as measured by W6/32 reactivity. B cell lines, peripheral blood lymphocytes, fibroblasts, a HeLa cell derivative, and HSB2, another T cell line, had equal amounts. Immunological cross-reactions between HLA-A, B, C antigens and beta 2-microglobulin and their homologues in other species were detected with the BBM.1 and W6/32 antibodies. The W6/32 antigenic determinant appears to be more highly conserved than that recognized by the BBM.1 antibody.


Idiotypic analysis of antibodies against the terpolymer L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT). IV. Induction of CGAT idiotype following immunization with various synthetic polymers containing glutamic acid and tyrosine. The immune responses of all inbred strains of mice specific to the synthetic terpolymer poly(LGlu60LAla30LTyr10), referred to as GAT10, are characterized by the presence of anti-GAT antibodies which share a common (CGAT) idiotype. In this report, we describe the ability of the synthetic polymers, LGlu33LAla33LTyr33, LGlu51-LAla34LTyr15 and poly-L(Tyr, Glu)-DLAla--LLys [(T,G)-A--L] to induce antibodies with CGAT idiotypic specificities. All of these polymers contain "GT"-related determinants. Following immunization with these polymers, antisera from responder mice bind the corresponding 125I-labeled antigen and 125I-labeled poly(LGlu50LTyr50) or GAT10. These antisera shared the CGAT idiotype which is associated with the antibody fraction with binding specificity for GAT10. Collectively, the present results indicate that GT-related determinants are required for the induction of the CGAT idiotype. Moreover, since the immune responses to these synthetic polymers are under distinct H-2-linked immune response (Ir) gene control, a mouse strain can be nonresponder to one polymer and responder to another; in this case, only the latter polymer induces CGAT idiotype. Thus, although the immune responses of inbred strains of mice to different polymers are under distinct Ir gene control, the antibody responses can be idiotypically related.


Antigen-specific suppressor factors produced by T cell hybridomas for delayed-type hypersensitivity. This study describes the generation of T hybridoma lines which secret factors specifically suppressing delayed-type hypersensitivity (DTH) to sheep red blood cells (SRBC). AKR strain-derived T lymphoma BW 5147 cells were fused with spleen cells from mice primed with SRBC and containing antigen-specific T suppressor cells for DTH. Supernants from the derived hybridomas were tested for suppression of either expression of induction of DTH to SRBC. Six lines produced specific suppressor activity for the expression of DTH; 4 lines produced suppressor activity for the induction of DTH of which only one line was antigen-specific. These lines were passaged in normal AKR mice, and the serum obtained had activity up to 10(-4) dilution. The factor was effective across the H-2 barrier.


Fine specificity of a continuously growing killer cell clone specific for H-Y antigen. H-Y-specific cytotoxic T cells were first cloned in soft agar and grown over a period of 8 months in media conditioned with supernatants from mouse and rat spleen cells stimulated with concanavalin A. The specificity of cloned cells and their cytolytic potential remained essentially unchanged over the entire culture period. In addition to lysing male target cells expressing H-2Db antigens, the cytolytic cells lysed also male as well as female cells expressing H-2Dd alloantigens. Seventeen out of eighteen subclones derived from the original clone revealed the same activity. The cells divide about every 17--20 h can be obtained in large quantities.


Adenosine triphosphate levels during anaphylactic histamine release in rat mast cells in vitro. Effects of glycolytic and respiratory inhibitors. The adenosine triphosphate (ATP) content of rat mast cells was studied during and after anaphylactic histamine release. The almost identical time course of ATP decrease from mast cells treated with either glycolytic or respiratory inhibitors supports the view that the ATP depletion was largely related to the histamine release process and not to an uncoupling of oxidative phosphorylation by an increased concentration of cytosol Ca2+. The ATP content of the cells was not restored within the 2 h of observation. No inhibition of lactate production from mast cells exposed to antigen in the presence of respiratory inhibitors and glucose was observed. Based on the lactate production from mast cells, the turnover time of ATP was calculated to be about 3/4 min.


Studies on the cytochemical localization of adenylate-cyclase activity in Dugesia lugubris s.I. The localization of adenylate-cyclase activity in Dugesia lugubris s.1. has been investigated cytochemically using 5'-adenylyl-imidodiphosphate as substrate. The enzyme was localized in mucous gland cells, in rhabdite cells, in intercellular spaces and also in nerve endings of this planarian. The presence of adenylate-cyclase on the membrane suggests that it might mediate different stimulus-secretion coupling by increasing cyclic AMP synthesis in specialized areas of the planarian.


[Enzymatic N-acetylation of tryptamine by brain homogenates of Locusta migratoria before and after intoxication by chlordimeform or lindane (author's transl)]. Brain homogenates of Locusta migratoria are found to possess enzyme capable of catalyzing the N-acetylation of tryptamine. A main product of the enzymatic reaction is isolated and identified as N-acetyltryptamine by chromatography analyse. Both insecticides, chlordimeform and lindane, inhibit enzyme activity. A direct correlation between the degree of intoxication and acetylation of tryptamine is described.


Antigenic correlation between rat brain synaptic vesicles and rat bone marrow B lymphocytes. Rabbit anti-rat brain synaptic vesicle serum reacted with thymocytes and B lymphocytes in cytotoxicity and immunofluorescence assays. Quantitative absorption analysis revealed that this antiserum contained antibodies specific for antigenic determinants on the surface membrane of a subpopulation of rat bone marrow B lymphocytes.


Measurement of human proinsulin by an indirect two-site immunoradiometric assay. An indirect two-site immunoradiometric assay is described for the measurement of human proinsulin in plasma. Polyethylene tubes coated with purified guinea-pig antibodies to insulin were used to extract proinsulin and insulin from plasma. Rabbit antibody to human C peptide was then added to react with the C-peptide moiety of the bound proinsulin. The uptake of this antibody was measured by the subsequent binding of 125I-sheep antibody to rabbit IgG. The binding of radioactivity to the tubes was a function of the proinsulin concentration in the sample. The sensitivity of the assay was 0.006 pmol/ml. Only 200 microliters of plasma was required in the assay and the 125I-labelled antibody was produced from readily available reagents. The polyethylene tubes remained stable for at least 5 months after coating. The mean fasting proinsulin level was 0.009 pmol/ml in sixteen normal subjects and 0.025 pmol/ml in twelve maturity onset diabetics. Oral glucose produced an 8 fold increase in proinsulin concentration but a decline in the plasma proinsulin/insulin molar ratio. Four patients with insulinoma had extremely elevated proinsulin levels and proinsulin/insulin ratios.


Isolation of histamine-containing cells from canine fundic mucosa. The cells of the fundic portion of canine gastric mucosa were dispersed by collagenase digestion and separated into fractions by sequential use of velocity sedimentation in an elutriator rotor followed by a density gradient separation. There was a close correlation between histamine content and number of mast cells in the different cell fractions. The mast cells possessed characteristic dense granules, which stained metachromatically, but did not release histamine on exposure to Compound 48/80. The most highly purified fractions contained 80% mast cells and a histamine content of 2.5 pg/mast cell.


Molecular hybridization probes for research in liver disease: studies with albumin cDNA. A major avenue of our research has been to develop molecular probes (cDNAs) for studying the pathogenesis of liver disease (i.e., applied molecular pathophysiology). During the last 2-3 yr, we have developed and used molecular hybridization to study regulation of albumin synthesis in normal, protein-calorie deprived, uremic, and cirrhotic rat liver. Our current work is directed toward cloning the albumin gene to permit further analysis of albumin transcriptional and posttranscriptional control. Molecular hybridization, DNA cloning, related techniques can be utilized to study the function of virtually any gene. Incorporation of such advances in basic research into meaningful studies of liver disease is an exciting challenge to modern academic hepatology.


[Human chromosome polymorphism and disordered reproductive function. II. C-variant chromosomes]. C-stained polymorphic variants of chromosomes 1, 9, 13--16, 21, 22 and Y were studied in married couples with reproductive failure (200 individuals) and in control couples having normal children and no spontaneous abortions and stillbirths. Location of heterochromatic segments, their size and heteromorphism of homologues were estimated. The individuals with reproductive failure were carriers of variants of chromosomes 9 and acrocentrics with higher content of heterochromatic material as well as with heterochromatic chromosome 9 significantly more frequently as compared with control individuals.


[Review. Clinical importance of the lymphocyte differentiation in peripheral blood]. The study of the lymphocyte subpopulations appears to be of diagnostic importance in immunodeficiencies and lymphatic disorders. In autoimmune diseases and cancer as well as in various other disease states changes of T and B lymphocytes are detectable, findings that can be of prognostic and therapeutic importance.


Comparative ultrastructural morphometrical studies in acute leukemia. Four hundred paramyeloblasts (from meyloblastic, promyelocytic, monoblastic and lymphoblastic types), isolated from peripheral blood of untreated leukemia patients, were studied by planimetric ultrastructural morphometry. The data of 19 parameters for these four paramyeloblastic types were compared with statistical methods. More central "scattered" heterochromatin was found from this morphometric investigation, i.e. early prophases in the lymphoblastic type of acute leukemia (these cases are more sensitive to therapy). The absolute mean values of the areas of whole cells, areas of the nucleus and nucleolus, areas of the heterochromatin and other indices show that different cell clones will undergo leukaemic transformation.


[Comparison of the cytomorphological classification with the cytochemical differentiation of acute lymphatic leukemia in children]. In 100 children with acute lymphatic leukaemia the cytomorphological subclassification of the pathological cell type was made according to Mathé and the French-American-British Co-operative group (FAB). In addition, all cases of leukaemia were differentiated according to their cytochemical type. Lymphoblasts from 10 cases of leukaemia could be subclassified immunologically. From 71 children will ALL the survival rates of those cases of leukaemia subclassified cytomorphologically and the cytochemical reactions were compiled and partially compared. Microlymphoblastic leukaemia could be found to be the most frequent type of ALL at children's age. Prolymphocytic leukaemias were characterized by a favourable survival rate and the highest percentage of ALL with the PAS type. Macrolymphoblastic and microlymphoblastic cases of leukaemia revealed no essential differences of survival rate, but significant differences of cytochemical reactions.


[Leukemic chromosome clone. I. Risk factor in the acute lymphatic leukemia of children]. As long as the aetiology of acute lymphatic leukaemia of children is not known its therapy is based on clinical experience. Among the values of experience those factors will play a part, the evidence of which during the ALL initial stage will be a risk for successful therapy and survival rate. This results in a choice of more aggressive variants of modern therapy schemes. In a cytogenetic study made in 35 children with ALL it was tested, whether even leukaemic chromosome clones will be a risk for the course of acute leukaemia. The duration of the first remission and survival rate were considered as criteria. The evidence of a leukaemic chromosome clone could be shown to be followed by a short survival rate, irrespective of the stage of the disease where the clone had been observed first. Thus, cytostatic therapy in those ALL patients who are affected with luekaemic chromosome aberration of stem line character should be aimed at the complete annihilation of the clone, irrespective of other remission criteria. The failure of blood and bone-marrow cultures as early as during the untreated initial stage indicated a primary cellular immuno-insufficiency. This combination of cell immuno-depression with high peripheral leukocytes connts and a primary mediastinal tumour or a generalizing lymphosarcoma respectively, was the highest risk up till now for the course of the disease. Judging from the duration of the first remission and the survival rates, the consecutive schemes of therapy did not differ in their effect on leukaemia with pathological stem lines. On the basis of the present study the impression could not be excluded that up till now long term survival rates could be attributed rather to individual manners of response than to the modern therapy scheme.


[Quantitative behavior of B-lymphocytes in infection susceptible children following a single therapeutic substitution with human gamma globulin]. In 15 children with a lowered gammaglobulin level a single substitution with HGG was made, with the impact of this substitution on the number of B-cells in the peripheral blood being examined by means of the direct fluorescence antibody technique. In 9 from 15 children the substitution had no influence on the number of B-cells. However, a significant increase of the number of B-cells could be observed in 6 from 15 children. 14 days after the substitution the number of B-cells lay within the normal range again.


Cytochemistry of nucleoproteids and some cathionic proteins in the peripheral blood leukocytes of patients with lung cancer. In 40 patients with untreated lung cancer cytochemical studies of the peripheral blood leukocytes were conducted by means of a cytological method for the simultaneous staining of nucleoproteids (RNP and DNP) and some cathionic proteins (after Zvetkova and Zvetkov [60]). Changes were detected in the RNP cytoplasmic contents of lymphocytes, of which the most outstanding were the reduction and uneven distribution of RNP granules, their frequent extracellular expulsion by means of microclasmatoses, as well as changes in the staining of cathionic proteins of RNP accompanied by an increased nuclear chromatin condensation in the small and medium-sized lymphocytes. Parellel to reducing of the percentage of these cells in the peripheral blood of patients with advanced neoplastic disease an increased number of lymphoblastoid and monoblastoid cells is established with RNP diffusely stained, but reduced in quantity and localized in the cytoplasmic periphery and projections (compared to Downey type II atypical cells). By means of one of the variants of the method (modified type of Feulgen's reaction) a characteristic distribution and structuring of the nuclear chromatin is established in mono- and polymorphonuclear cells, most clearly expressed in the nuclei of monocytes and monoblastoid cells, as well as in nuclei of neutrophil granulocytes. In these cellular types a more specific nuclear modelling (microhypersegmentation) is observed resulting in multiple irregular nuclear projections on the nuclear surface, probably caused by subkaryolemal distribution of uneven chromatin thickenings. The changes are also recorded in the cathionic protein containing secondary cytoplasmic granules in granulocytes-neutrophils and eosinophils, probably associated with changes in the lysosomal and phagocytic functions of these cells in neoplastic diseases. The authors discuss the importance of the obtained results in connection with data on the participation of lymphocytes and neutrophils in the immune response to tumour antigenic stimuli during the course of the neoplastic process, as well as with data on the suppressive effect of antigenic (serum, viral) factors, possibly affecting the synthesis and the transport of cellular nucleoproteids (RNP and DNP) in leukocytes of cancer patients.


The effect of intravenous injection of ethyl palmitate emulsion on sequestration of thermally damaged erythrocytes in rats with experimentally induced hypersplenism, and in normal rats. The authors attempted at experimental elimination of sequestration function of the spleen in Wistar rats using an i.v. injection of ethyl palmitate emulsion, both in "hypersplenic" animals being long-term applied i.p. methyl cellulose solution, and in control rats. In the rats clearance of 51Cr-labelled and thermally damaged erythrocytes from blood was examined and their sequestration in the spleen and liver followed. The ethyl palmitate injection resulted in both experimental groups in a significant decrease of the erythrocyte counts sequestrated in the spleen, and significant prolongation of the elimination half time for thermally damaged erythrocytes from the blood.


[Frequency of anemia in Rostock students]. For the purpose of determining the frequency of anaemia a check-up was made in 735 students (498 females, 237 males) at an age of at least 19 years at the beginning of their university studies. 9.4% of them had subnormal haemoglobin values. 91.3% of them were female students. A comparison between the test persons with a decrease of haemoglobin values and those without it on the one hand and data gathered from the students' health certificate on the other hand, revealed no ensured aetiologic connection. Analyzing the state of health of that group with lowered Hb-values showed that more than half of them suffered from diseases (mostly infections) at the time, when the blood examination was made. A check-up made one year later resulted in a high percentage of normalized values. Comparisons taken from medical literature clearly indicate that the frequency of anaemia of this student population examined here is relatively low in total. From an organizational point of view, complete check-up examinations shoudl be carried out or induced for female students by the outpatient departments working at the institutions of higher learning because they provide better possibilities of covering them.


[Case contribution to the study of hemoglobin C disease]. Haemoglobin-C disease represents a haemoglobinopathy seldom occurring in our latitudes. In its homocygotic form it coincides with haemolytic anaemia and splenomegaly. The diagnosis of haemoglobin-C disease could be ensured in a 33 years old Yugoslav patient who had been treated because of being suspected of a venous thrombosis of the spleen. The knowledge of this disease taking prognostically a benign course seems to be important in order to prevent patients from unnescessary diagnostic and therapeutic measures.


[Participation of the hypophyseal-adrenal cortex system in thrombin clearance during immobilization stress]. The examination carried out with thrombin marked by 131J resulted in a considerable increase of the thrombin clearance rate in healty male rats during the stress (caused by an immobilization lasting 30 minutes) and in an increase of thrombin deposits in the liver. A further increase of thrombin clearance occurred by the combination of immobilization and administration of ACTH. Contrary to ACTH the thrombin clearance is not stimulated in healthy animals by hydrocortisone. Thrombin clearance and thrombin deposits in the liver are lowered in adrenalectomized rats. In these animals the administration of ACTH does not result in an increase of thrombin clearance. The rate of thrombin clearance is normalized in adrenalectomized animals after administering hydrocortisone without as well as under conditions of stress. In adrenalectomized animals having received hydrocortisone as well as in healthy animals the administration of ACTH will results in an increase of thrombin clearance. From these experiments the conclusion can be drawn that ACTH will increase the intensity of thrombin clearance in stress and that hydrocortisone plays a transmitting part here.


[Immunological concentration determinations of the streptokinase preparation Awelysin dissolved in various infusion solutions for its stability evaluation]. By means of the electro-immuno-assay according to Laurell the decrease of concentration was determined for the streptokinase preparation Awelysin dissolved in various infusion media at 20 degrees C. The stability behaviour of a streptokinase solution of 1.000 IE/ml showed a significant decrease of concentration after 24 hours (alpha less than 5%) as compared with the initial concentration. However, in comparison no significant differences as to the streptokinase stability could be found by using different infusion solutions. At 20 degrees C the decrease of streptokinase concentration may be neglected up to 24 hours in an infusion solution of streptokinase previously prepared.


Platelet fibrinogen: subcellular localization by means of immunofluorescent studies in normals and in congenital afibrinogenemia. We have studied the site of fibrinogen localization in normal platelets and in the platelets of a patient with congenital afibrinogenemia (CA). The methods employed were: direct immunofluorescence technique (DIT) and indirect immunofluorescence technique (IIT). By means of the DIT normal platelets were shown to have a clear peripheral staining. Such staining disappeared after treatment with proteolytic enzymes and after specific blocking experiments. Such peripheral staining of platelets was absent in congenital afibrinogenemia even after fibrinogen infusion. By means of the IIT platelets were shown to have a considerable amount of fibrinogen. Such protein was demonstrated to represent an important part of platelet surface, since intact platelets were able to absorb completely a specific antifibrinogen antiserum.


[Drug-induced lung diseases].

[Drug-induced lung diseases]. Drug-induced lung diseases may present themselves as bronchial reactions (e.g. bronchial asthma), diseases of the parenchyma (e.g. pulmonary infiltrates with eosinophilia, diffuse fibrosing alveolitis), of the pulmonary vasculature (vasculitis) and of the pleura (e.g. pleurisy or pleural fibrosis). Pathogenetically the two most pertinent types of reaction are hypersensitivity or toxic reactions, and less often biologic reactions such as opportunistic infections after cytotoxic and immunosuppressive therapy. Many drug-induced respiratory diseases are reversible upon withdrawal of the offending agent; others may be irreversible or even progress.


[Clinical valence of alpha-fetoprotein (author's transl)]. Among carcinoembryonic antigens CEA and AFP have an exceptional range. The detection of AFP in the serum is a valuable and sensitive serological marker of primary hepatic carcinoma. The quota of primary hepatic carcinoma without AFP-synthesis is for less than 5%.


Purification of the subunit Clq from the first component of equine complement. Initial separation and concentration of Clq from fresh, normal equine serum was accomplished by precipitation in 0.02 M acetate buffer, pH 5.5, at 4 degrees for 24 h. The re-dissolved precipitate was clarified by centrifugation at 80,000 g for 1 h and then dialysed against Tris-HCl buffer (0.05 M, pH 8.0) containing 10-3 M EDTA. The clarified dialysate remained biologically active at 5 degrees for at least 4 weeks. Biological activity of equine Clq was determined by assay of its ability to agglutinate sensitized sheep erythrocytes (EA). Following ammonium sulphate fractionation, Sepharose 4B gel filtration yielded three major peaks. Two protein bands were demonstrated on analysis of the second Sepharose peak by disc acrylamide electrophoresis, pH 8.3. Elution of the protein bands showed EA-agglutinating activity only in the band which migrated furthest toward the cathode. Equine Clq isolated by this method yielded an approximate forty-fold purification in specific activity. Some properties of equine Clq were characterized. Equine Clq was heat-labile, as shown by loss of its EA-agglutinating activity after heating 58 degrees for 15 min. Moreover, storage at 4 degrees and freeze-thaw cycles greatly reduced EA agglutination. Preliminary determination of the sedimentation coefficient indicated that it was comparable to that reported for human and rabbit Clq.


Studies on immune tolerance to heat-aggregated human IgG in rabbits: its relevance to the production of rheumatoid factors. Rabbits neonatally tolerized with native monomeric human IgG and subsequently immunized with heat-aggregated human IgG emulsified in Freund's complete adjuvant, at a time when B cells but not T cells should have spontaneously recovered from tolerization, resulted in the production of an antibody resembling human rheumatoid factors directed against a single antigenic determinant in the Fc region of the IgG molecule. Rabbits immunized in an identical fashion after adult tolerization treatments failed to respond during the period corresponding to B-cell tolerance, but subsequently produced antibody or similar specificity to that produced by neonatally tolerized animals. These results indicate that no completely new antigenic determinants are created on heat aggregation of IgG, but there appears to be some potentiation of immunodominance of an already existing determinant. A possible mechanism for the production of rheumatoid factors in rheumatoid arthritis is suggested.


Regulatory role of suppressor T cells in the expression of delayed-type hypersensitivity in mice. I. Transient appearance of suppressor T cells for the expression of delayed footpad reaction induced with lipid-conjugated lysozyme. Delayed footpad reaction (FPR) to lysozyme (Lys) in mice was induced without antibody responses by lipid-conjugated lysozyme (D.Lys). This FPR was suppressed by priming s.c. with a high dose (10 mg) of Lys 2 weeks previously (unresponsiveness). Spleen cells from the unresponsive mice suppressed antigen-specifically FPR in mice previously immunized with D.Lys, and also suppressed passive transfer of FPR by D.Lys-immune lymphoid cells into normal mice. The suppressive activity of the spleen cells was abolished by treatment with anti-phi anti-serum and complement. The suppressor cells occurred also in the thymus of unresponsive mice. Unresponsiveness was induced in mice immediately after priming with Lys and persisted at least up to 7 weeks after the induction. In contrast, suppressor cells appeared only 2 weeks after induction of unresponsiveness in both the spleen and the thymus but were no longer detectable 3-7 weeks later, although donor mice remained fully unresponsive. These results suggest that antigen-specific suppressor T cells are involved in the regulation of the expression of FPR only for a definite period of time in unresponsive mice.


Regulatory role of suppressor T cells in the expression of delayed-type hypersensitivity in mice. II. Soluble factor from thymic suppressor cells stimulated with antigen in vitro and its possible interaction with macrophages. Thymus cells from mice primed s.c. with a high dose (10 mg) of lysozyme (Lys) specifically suppressed delayed footpad reaction (FPR) in mice previously immuned with lipid-conjugated lysozyme (D.Lys), and also suppressed the transfer of FPR by D.Lys-immune spleen cells into normal mice. Furthermore, they inhibited antigen-stimulated DNA synthesis of D.Lys-immune spleen cells in vitro. If the suppressor thymus cells were cultured with Lys in vitro, they produced soluble factor which depressed the ability of D.Lys-immune spleen cells to transfer FPR. Both supernatant of culture without Lys and extract of suppressor thymus cells were inactive in supression of FPR. The suppressor factor was antigen-specific because its suppressive activity was absorbed with Lys but not with an unrelated antigen lactalbumin. The factor failed to depress the ability of D.Lys-immune spleen cells to transfer FPR when the spleen cells were depleted of glass-adherent cells. In addition, incubation of peritoneal exudate cells from normal mice with the factor rendered the cells suppressive for passive transfer of FPR. These results suggest that the suppressor factor depresses the effector function of T cells responsible for FPR possibly via macrophage.


Thymus dependence of the IgG response: role of T cells is restricted to non-specific rather than antigen-specific factors. We have been studying the requirement for T cells in the proliferation and maturation of partially differentiated memory cells into IgG antibody secreting cells. The role of T cells in such a system can be replaced by non-antigen-specific T-cell factor(s) present in the supernatant of a 24 h mixed lymphocyte reaction (TRF) but not by antigen-specific factors released upon in vitro induction of T-helper cells with antigen. A combination of antigen-specific and non-specific factors showed no evidence of synergy between the two sets of factors and we therefore conclude that the antigen-specific factors tested play no role in the later stages of B-memory cell maturation to IgG-antibody secreting cells.


An absolute requirement for 2-mercaptoethanol in the in vitro primary immune response in the absence of serum. Using a serum-free medium, high recoveries of plaque-forming cells (PFCs) were obtained up to 5 days of incubation in  the in vitro primary immune response. Under these conditions, the presence in the medium of 2-mercaptoethanol was an absolute requirement. In order to obtain maximal recoveries of PFCs, the antigen and 2-mercaptoethanol had to be present from the start of the culture, while the addition of fetuin could be delayed for 24 h without any loss of PFCs. The 2-mercaptoethanol could be replaced by (dialysed) human serum.


Amplification of plaque-forming cells in the spleen after intracloacal antigen stimulation in neonatal chicken. Neonatal chickens were primed by multiple or a single injection with sheep red blood cells (SRBC) into the cloacal lumen and then  were challenged by intravenous injection with the same antigen. Direct plaque-forming cells (PFC) in the spleen were markedly increased shortly after intracloacal priming. This enhancing effect was antigen specific and was abolished by surgical bursectomy 24 h after intracloacal priming. A similar effect was also found in mature adult chickens, although the degree of enhancement was far less than that found in neonatal chickens. These results suggest that a certain stage of B-cell differentiation in the bursa of Fabricius (BF) is regulated by antigenic stimulation.


Purification and immunobiological properties of R antigen and its relation to M protein of type 3 group A Streptococcus. R protein was extracted from type 3 group A streptococci with hot (95 degrees C) HCl and was purified by ammonium sulfate precipitation followed by molecular-sieve and ion exchange chromatography. Although the R3 antigen was present in a heterogeneous population of proteins ranging from 78,000 to 100,000 daltons in size, we were able to separate an R-rich fraction that contained minimal amounts of heterogeneous proteins as indicated by electrophoresis in sodium dodecyl sulfate-polyacrylamide gels. The final yield of the purified R protein was approximately 15 mug (dry weight) per g (wet weight) of washed and sedimented streptococci. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated a molecular size of approximately 78,000 daltons. Amino acid analysis showed lysine, glutamic acid, alanine, and aspartic acid as the predominant amino acids. A detailed comparison of the purified R3 protein with type 3 M protein indicated a similarity in composition and order of frequency of amino acids. However, the R3 antigen was found to be distinctive from the M3 antigen in agar gel diffusion tests. In addition, R3 and M3 proteins behaved differently in opsonophagocytosis tests and opsonization inhibition tests. Thus, R3 and M3 proteins produced precipitin lines of nonidentity with an unabsorbed antiserum against whole type 3 streptococci: M3-specific antiserum, but not R3-specific antiserum, enhanced the phagocytosis of type 3 streptococci. Purified M3 but not R3 protein was capable of inhibiting the type-specific opsonization of type 3 streptococci. The physicochemical resemblance between M and R proteins in general suggests a common genetic origin. Perhaps R proteins are variant forms of M proteins from which the antiopsonic determinant has been deleted.


Immunological specificity of natural opsonins and their role in the cross-reactivity between Staphylococcus aureus Mardi and Escherichia coli 101. The immunochemical specificity of the observed cross-reactivity between Escherichia coli strain 101 and Staphylococcus aureus strain Mardi was examined. The cross-reactivity was shown to be dependent upon mucopeptide antibodies which are present in normal and immune sera. Although both organisms contained surface antigens with immunodominant glucuronic acid residues, in vitro phagocytosis studies indicated that antibodies directed against these antigens were not significantly involved in the opsonization process. Rather, antibodies with mucopeptide specificity were shown to be involved in the in vitro phagocytosis of these organisms by polymorphonuclear leukocytes. The mucopeptide antibodies, which were found in both nonimmune and immune sera, were shown to be effective in opsonizing both the S. aureus strain and the E. coli strain. The ubiquitous distribution of E. coli strains containing mucopeptide antigens common to most bacteria suggests that these organisms may be responsible for the wide prevalence of natural staphylococcal opsonins with mucopeptide specificity in normal sera.


Behavior of a surface antigenic determinant of lateral flagella of Vibrio parahaemolyticus. The behavior of a surface antigenic determinant (SA) of lateral flagella of Vibrio parahaemolyticus was studied. SA, located on the surface of flagellar filaments, is responsible for H-agglutination and specific for serotype. Results of H-agglutination inhibition tests demonstrated that SA could not be detected on the flagellin molecule when the flagellar filaments were dissociated to flagellin monomers by heating or treatment with urea, sodium dodecyl sulfate, HCl, or acetone, although SA could be detected on short flagellar fragments obtained by milder heat treatment. When flagellar filaments were reconstituted from flagellin monomers, SA was detected on the surface of the filaments. These results suggest that SA is buried in the flagellin molecule with dissociation of flagellar filaments to flagellin monomers or steric configuration of SA itself is altered to a different form which cannot react with the responsible antibody.


The use of hapten-polysaccharide conjugates for the induction of B-cell tolerance involving IgE responses. III. Specific tolerance induced by sulphonamide-substituted levan in the guinea pig. Inbred strain 2 and (strain 2 x strain 13)F1 guinea pigs were made allergic by intraperitoneal injections of sulphonamide-chicken gamma-globulin (CGG) conjugates. Two sulphonamides were used: 4-sulphanilamide benzoic acid (4-SABA) and sulphamethoxazole (SMX). 4-SABA was coupled to CGG through its carboxylic group and SMX was coupled following diazotization of its sulphanilamide group. The anti-4-SABA and anti-SMX reaginic antibodies formed did not show any cross-reactivity with each other. Injection of 4-SABA coupled to native levan effectively suppressed the allergic responses of these guinea pigs when given either prior to or after immunization with 4-SABA-CGG. This treatment is specific as it did not affect anti-SMX or anti-CGG reaginic responses. Guinea pigs seem more sensitive to the regime used for tolerance induction than correspondingly sensitized mice in that less tolerogen is required on a body weight basis.


Effect of cyclic AMP on histamine release induced by compound 48/80. Bu2 cAMP(N6, O2'-dibutyryl adenosine-3',5'-cyclic monophosphate) inhibited the response of rat peritoneal mast cells to compound 48/80 in the presence of calcium ions. In the absence of calcium, the nucleotide partially prevented the desensitization induced by chelating agents. The response of cells, allowed to accumulate Bu2 cAMP in the presence of calcium (to avoid depletion of intracellular stores of the ion) and then challenged in the absence of extracellular calcium, was also inhibited. These results are discussed in terms of the postulated effects of Bu2 cAMP on the calcium-gatubg mechanism operative in histamine secretion.


In vitro release of eosinophil chemotactic factor of anaphylaxis (ECF-A) from guinea pig skin. An in vitro release of the eosinophil chemotactic factor of anaphylaxis (ECF-A) in anaphylactic reactions was studied using skin slices from actively sensitized guinea pigs. ECF-A and histamine were released in vitro from actively sensitized guinea pig skin by antigen challenge. The magnitude of ECF-A release did not always parallel that of anaphylactic histamine release in guinea pig skin, suggesting that there may be no correlation between the amount of ECF-A and that of histamine released from the skin in anaphylactic reactions in guinea pigs.


Fragility of attenuated Rauscher leukemia virus. Some biochemical properties of virulent (RRL+) and attenuated (RCL-) Rauscher leukemia virus were compared. It was shown that the reverse transcriptase (RT) activity of 'aged' as well as 'fresh' RCL- virions was 25--30% of that found for RRL+. The thermal sensitivity of the RT was the same for both viruses. A 60--70S RNA could be extracted from 'aged' RRL+, while no high molecular weight RNA was obtained from 'aged' RCL-. After centrifuging in sucrose gradient, most of the RT activity and 3H-labeled RNA of 'aged' RRL+ was recovered at 1.14--1.16 g/cm3, while for 'aged' RCL- no labeled RNA, and at most 10% of the original RT activity were found in the same zone. The fragility seemed to increase in the course of aging, since 'fresh' RCL- banded at 1.14--1.16 g/cm3 as did 'fresh' RRL+. Also, 3H-labeled viral RNA was found in the viral bands, from which 60--70S RNA could be extracted. Molecular hybridizations showed that 20% of the nucleic acid sequences related to Rauscher leukemia virus found in the RNA of RRL+-infected cells were missing in the RNA of RCL--infected cells.


Monkeypox virus as a source of whitepox viruses. Monkeypox virus cloning and isolation of the so-called 'white' clones from white pocks which this virus forms on the chorioallantoic membrane (CAM) were carried out. The isolated clones were stable and differed considerably from the parental strain. By their properties, they were identical to whitepox viruses formerly isolated from wildlife monkeys and rodents in Equatorial Africa. Besides stable 'white' clones, a number of virus cultures in the process of cloning were obtained which differed in quantitative content of virions, forming on CAM white pocks and pocks with hemorrhages. It appeared that the properties of the viral population as a whole (reaction type on rabbit skin, hemagglutination activity, etc.) depended on the rate of virions produced with different characteristics.


Lysosome stability during lytic infection by simian virus 40. By 48 h postinfection, 40--80% of SV40-infected CV-1 cells have undergone irreversible injury as indicated by trypan blue staining. Nevertheless, at this time the lysosomes of these cells appear as discrete structures after vital staining with either acridine orange or neutral red. Lysosomes, vitally stained with neutral red at 24 h postinfection, were still intact in cells stained with trypan blue at 48 h. Acid phosphatase activity is localized in discrete cytoplasmic particles at 48 h, as indicated by histochemical staining of both fixed and unfixed cells.


Chemotherapy of head and neck cancer.

Chemotherapy of head and neck cancer. Chemotherapy has traditionally been relegated to a late palliative role in the management of head and neck cancer. For advanced disease in patients previously treated with surgery or radiation therapy, methotrexate, which has a response rate of approximately 50%, is the best single agent available. Other orally or systemically administered single agents--such as bleomycin, adriamycin, hydroxyurea, and cis-platinum--produce responses less frequently than methotrexate. Intraarterial chemotherapy with single-agent methotrexate or with combinations--including methotrexate, vinblastine sulfate (Velban), cyclophosphamide (Cytoxan), and 5-fluorouracil (5-FU)--can match the response rate of systemic methotrexate, but all require inpatient treatment and significant technical expertise. Programs utilizing chemotherapeutic combinations have not produced remission rates or durations of remission greater than those achieved with methotrexate alone. Chemotherapy has also been used in combination with other modalities. In two studies, chemotherapy combined with irradiation produced improvement in median or overall survival in comparison to irradiation alone. Many similar studies have been negative. Recent trials of chemotherapy used preoperatively in the treatment of head and neck cancer have not yielded decreased recurrence rates or increased overall survival rates in comparison to surgery alone, but they do appear to have prolonged the disease-free interval following definitive surgery.


Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Sirius Red, a strong anionic dye, stains collagen by reacting, via its sulphonic acid groups, with basic groups present in the collagen molecule. The elongated dye molecules are attached to the collagen fibre in such a way that their long axes are parallel. This parallel relationship between dye and collagen results in an enhanced birefringency. Examination of tissue sections from 15 species of vertebrates suggests that staining with Sirius Red, when combined with enhancement of birefringency, may be considered specific for collagen. An improved and modified method of staining with Sirius Red is presented.


The use of peroxidase-labelled Limulus polyphemus agglutinin for the histochemistry of sialic acid-containing glycoproteins in light microscopy. The usefulness of a lectin, Limulus polyphemus agglutinin (LPA) has been tested in a series of mammalian tissues with sialic acid-containing glycoproteins. In nearly all the tissues employed, the positive peroxidase-labelled LPA diaminobenzidine (LPA-PO-DAB) reaction of various histological structures was markedly diminished in intensity or abolished, following digestion with neuraminidase. In the same tissues, sialic acid added with LPA-PO abolished the LPA-PO-DAB reaction or notably suppressed its intensity. In the majority of the tissues tested, the LPA-PO-DAB-Alcian Blue (AB) (pH 1.0 or 2.5) procedures appear to be useful dual staining methods which enable one to colour selectively sialic acid-containing and other acidic carbohydrates. In view of the endogenous peroxidase activity in particular histological structures, however, appropriate control staining procedures should be performed when the LPA-PO-DAB procedure is employed, either alone or in combination with AB procedures, to determine the histochemical properties of sialic acid-containing glycoproteins.


Evaluation of the glyoxal-bis-(2-hydroxyanil)-method for staining of calcium in model gelatin films and pancreatic islets. The nature of tissue calcium, detectable with glyoxal-bis-(2-hydroxyanil), (GBHA), was investigated using gelatin films as model. The results indicate that in the films the procedure detects only the calcium fraction which was ionized in the original gelatin solution. The GBHA staining intensity (absorbance) appeared to be linear with the amount of ionized calcium in the range from 0 to 2 micrograms/cm2. The method allows detection of amounts of ionized calcium as low as 0.15 micrograms/cm2 or 0.0015 pg/mu2. For the measurement of calcium in pancreatic tissue of fed rats, the tissue was subjected to freeze-substitution at -80 degree C in acetone containing 1% oxalic acid. Adjacent sections were stained with either GBHA or aldehyde-fuchsin (AF). Exocrine tissue hardly stained with GBHA whereas islet tissue stained intensely. For GBHA as well as for AF a variation in staining intensity (visual evaluation) between islets was observed. Islet GBHA- and AF-staining intensities did not correlate. The AF-staining intensity but not the GBHA-staining intensity decreased with increasing islet diameter. Also in pancreatic islet tissue the GBHA method appears to be very sensitive and reproducible and small differences in islet GBHA-staining intensity can be detected. The results indicate that between islets differences in ionized calcium content exist. These differences do not correlate with the degree of B-cell granulation.