Modulation of alphafetoprotein synthesis in the early postimplantation mouse embryo. The visceral endoderm of mouse egg cylinders on the 7th and 8th days of gestation is divided into the visceral embryonic (VE) endoderm cell population which synthesizes alphafetoprotein (AFP), and the visceral extra-embryonic (VEX) endoderm population which does not synthesize AFP. Embryonic (E) and extra-embryonic (EX) ectoderm and visceral endoderm tissues were enzymically separated, reassociated in different combinations, and cultured in vitro for 48 h. The immunoperoxidase reaction on sections of cultured tissues showed that both VE and VEX endoderm cells synthesize high levels of AFP when cultured in isolation or in association with E ectoderm, but do not synthesize AFP when in close association with EX ectoderm. Both 7th and 8th day VEX endoderm cells synthesize detectable levels of AFP 12 h after isolation, and contain high levels by 24 h. It is concluded that both VE and VEX endoderm cells have the ability to synthesize AFP, but modulation of expression occurs through an inhibitory influence of the EX ectoderm.


Cellular and genetic restrictions in the immunoregulatory activity of alpha-fetoprotein. II. Alpha-fetoprotein-induced suppression of cytotoxic T lymphocyte development. Alpha-fetoprotein (AFP), a major component of fetal and newborn sera, was shown to exert significant immunosuppressive activity on the in vitro generation of cytotoxic T lymphocytes (CTLs). This suppression proved independent of the suppressibility of the mixed leukocyte culture activation phase, since strain combinations whose proliferative responses were refractive to AFP-induced suppression also failed to develop demonstrable CTLs in the presence of AFP. Several strain combinations were also found in which normal generation of CTLs occurred in cultures containing AFP. This refractive nature correlated with the presence of nonsuppressible lymphocyte-stimulating alloantigenic systems on the stimulating cell population. These data provide the basis for proposing several possible mechanisms for AFP-induced suppression of T-cell-mediated cytotoxicity, as well as suggesting that the primary target of this suppression is the proliferating helper T cell precommited to respond towards the major histocompatibility complex-associated lymphocyte-activating determinants.


Induction and mode of action of suppressor cells generated against human gamma globulin. I. An immunologic unresponsive state devoid of demonstrable suppressor cells. A model of unresponsiveness to human gamma-globulin (HGG) which is maintained in the absence of demonstrable suppressor cells has been described. A/J mice were tolerized with deaggregated HGG purified from a variety of sources. The spleen cells from these tolerized mice were assessed for their ability to suppress the response of normal spleen cells to HGG when transferred into lethally irradiated mice. All of the HGG preparations obtained from commercial sources as Cohn fraction II of pooled, outdated plasma induced suppressor cells to HGG, although not of equal magnitude. However, suppressor cells could not be demonstrated in the spleens of mice tolerized with deaggregated HGG purified from the plasma of a healthy individual. This inability to detect suppression was independent of the method of purification of the HGG and of the time of assessment of the putative suppressor cells after tolerization. Similarly, deaggregated HGG isolated from an IgG1 lambda-myeloma protein induced unresponsiveness to HGG but did not stimulate demonstrable suppressor cells. These data suggest that suppressor T cells are not involved in the maintenance of tolerance to this antigen, although they may play a regulatory role in the immune response to HGG. Support for this concept was obtained by assessing the duration of unresponsiveness in the T and B lymphocytes of mice tolerized with the various HGG preparations. Mice tolerized with the HGG preparations that stimulated little or no suppression were among the last to recover responsiveness. Indeed, there was no consistent correlation between the level of suppressor cell activity and the degree of unresponsiveness in either the splenic T or B lymphocytes. Thus, although certain HGG preparations may provide a tool for the generation of antigen-specific suppressor T cells, the utilization of these suppressive preparations may be inappropriate for the investigation of the mechanisms of the induction and maintenance of the unresponsive state.


T-cell-specific murine Ia antigens: serology of I-J and I-E subregion specificities. (B10 X B10.D2)F1 mice were immunized with B10.A(5R) concanavalin A-stimulated thymocyte blasts. The genetic disparity between donor and recipient was restricted to the I-J and I-E subregions of the murine major histocompatibility (H-2) complex. A high-titered, T-cell-specific anti I-JkEk serum was obtained. The antiserum lysed 27-30% of haplotype k, q, or s lymph node cells, 5.3 +/- 2% of haplotype k spleen cells, and did not lyse thymocytes. Nylon wool-passed lymph node or spleen cells (H-2k) showed considerable reactivity with anti-I-JkEk serum (35-40% lysis); anti-Thy1.2 plus complement-treated spleen cells did not react (less than 5% lysis). I-Ek antibody was detected by B10.A(3R) lymph node cell reactivity (20% lysis), whereas reaction with H-2k lymph node cells after B10.A(3R) absorption demonstrated IJk antibody (12% lysis). Lymphocyte activation with alloantigen or mitogen led to increased anti-I-JkEk serum reactivity. These results, showing antibody production to at least two T-cell Ia antigenic determinants by concanavalin A thmocyte blast immunization, suggest that a group of I-region-encoded T-cell specificities may not have been detected using conventional immunization protocols because they would not have comprised a major antigenic component of the immunizing cell population. The existence of multiple Ia antigenic determinants unique to T lymphocytes would have important implications for serological and functional studies of T-cell subpopulations.


Spatial requirements between haptenic and carrier determinants for T-dependent antibody responses. To gauge the proximity between cooperating T and B cells required for effective triggering of antibody production, guinea pigs were immunized with bifunctional antigens in which the haptenic and carrier determinants were separated by rigid spacers of varied dimension. These took the form 2,4-dinitrophenol-(proline)n-L-tyrosine-p-azobenzenearsonate, where n varied from 1 to 40 proline residues. Animals immunized with n = 10 and n = 22 compounds made strong anti-DNP antibody responses, whereas animals immunized with bifunctional compounds containing longer spacers did not make antibody detectable by precipitation. It can be calculated on the basis of very strong physicochemical evidence for the rigidity and axial translation of poly-L-proline chains in solution that the cut-off point for effective interaction between T and B cells lies between 69 and 97 A U.


Shared antigenic determinants by mitogen receptors and antibody molecules to the same thymus-independent antigen. The antibody response to dextran B1355 is thymus independent, and in high responder mice, over 90% of the antibodies carry the idiotype of an alpha-1,3 binding myeloma protein (J558). The present experiments demonstrate: (a) dextran B1355 is a B-cell mitogen both in a strain which carries the J558 idiotype on antibodies and in a low-responder strain which does not express that idiotype on antibodies to dextran; (b) anti-idiotypic antibodies to J558 recognize a dextran-specific surface receptor on 10--15% of all splenic B cells in those two strains as well as in all strains so far tested; (c) as shown by inhibition experiments such surface receptors cross-react with J558, and (d) anti-idiotypic antibodies are mitogenic for spleen cells of both strains resulting in B-cell proliferation and maturation to polyclonal antibody secretion.


Purification of functional, determinant-specific, idiotype-bearing murine T cells. Strain A/J mice immunized with azobenzenearsonate (ABA)-mouse IgG conjugates develop suppression for anti-trinitrophenyl(TNP) responses to doubly conjugated (ABA,TNP) proteins. This suppression is specific for the ABA epitope and is mediated by T cells in cell transfer experiments. ABA-binding T cells from suppressed animals were purified by a two-stage procedure in which B cells were removed from spleen cell populations by adherence to plastic surfaces coated with anti-mouse Ig antibody, followed by binding the nonadherent population (more 95 percent Thy-1-positive) to surfaces coated with ABA-protein conjugates. Approximately 90 percent of the cells recovered by temperature-dependent elution from the ABA plates (similar to 2 percent of the spleen cells) bound antigen immediately afterward, and up to 50 percent of the cells bound anti-cross-reactive idiotype antibody. On the other hand, the nonadherent T-cell population was completely negative in the antigen- binding and idiotype assays. Another distinguishing feature of the two T-cell populations was that 78 percent of the adherent cells, but only 2 percent of the nonadherent cells, were Ia positive, although the specific I-region marker(s) expressed on the cells was not identified. The biological function of the antigen-binding T cells was investigated using a standard cell transfer protocol. Suppressor cells were enriched in the adherent population by a factor of at least 25, establishing that functional, epitope-specific, idiotype-bearing T cells can be significantly purified by this procedure. Note Added in Proof. We have recently isolated two types of ABA-binding molecules biosynthetically labeled with (35)S-methionine from NP-40 lysates of purified antigen-specific T cells. The molecules were purified by adsorption onto an ABA-Sepharose immunoadsorbent followed by elution with 9 M urea. Autoradiograms of SDS-PAGE of the eluates revealed components with tool wt of approximately 60,000 and 33,000 dahons. These molecules were not present in eluates from a bovine IgG-Sepharose control immunoadsorbent and thus represent specific ABA-binding products synthesized by T cells.


Immunological studies of aging. IV. The contribution of thymic involution to the immune deficiencies of aging mice and reversal with thymopoietin32-36. Aged mice preferentially lose the capacity to make IgG and high affinity PFC after immunization with the T-dependent antigen DNP-BGG. We have found that thymectomy accelerates the appearances of these immune deficiencies associated with aging. When splenocytes from old mice are transferred to young lethally irradiated, syngeneic mice and the recipients immunized 7 wk later, the number of IgG and high affinity PFC was increased compared to the response of old splenocytes transferred to young thymectomized mice. These immune deficiencies of aged mice were also reversed when old mice were treated with thymopoietin in vivo or splenocytes from old mice were incubated with thymopoietin before adoptive transfer to young irradiated, thymectomized syngeneic mice. The T-cell independent response to DNP-Ficoll was less impaired than the T-cell dependent response to DNP-BGG in old animals. These data suggest that a decline in thymic function that occurs during aging may contribute to the immunological deficiencies of old animals.


Expression of an onco-developmental antegen among avian species. Rabbit antisera directed against an onco-developmental antigen on chicken red blood cells have been serologically dissected through specific adsorptions. It is now possible to detect 13 antigenic determinants with the fractionated antisera. The onco-developmental antigen referred to as chicken fetal-leukemic antigen (CFA) is fetal-specific in the white Leghorn chicken, being present on the embryonic but not adult peripheral red blood cells of non-being present on the embryonic but not adult peripheral red blood cells of non-leukemic birds. However, one or more of the onco-developmental antigenic determinants have been detected on adult peripheral red blood cells of non-Gallus avian species, as well as on red blood cells from two adult chicken varieties. For phylogenetic purposes, red blood cells from avian species were characterized for their combinations of CFA determinants. Comparisons among species revealed specific patterns of antigenic expression within phylogenetic groups. Several CFA determinants were restricted in their occurrence to species within a single family, and one determinant was found in all cases where CFA was expressed. The distribution of CFA determinants was used to determine immunological distances among four Galliform species. These distances agreed with the immunological relationships established using different serological markers.


Interaction of ions and water in gramicidin A channels: streaming potentials across lipid bilayer membranes. For very narrow channels in which ions and water cannot overtake one another (single-file transport), electrokinetic measurements provide information about the number of water molecules within a channel. Gramicidin A is believed to form such narrow channels in lipid bilayer membranes. In 0.01 and 0.1 M solutions of CsCl, KCL, and NaCl, streaming potentials of 3.0 mV per osmolal osmotic pressure difference (created by urea, glycerol, or glucose) appear across gramicidin A-treated membranes. This implies that there are six to seven water molecules within a gramicidin channel. Electroosmotic experiments, in which the water flux assoicated with current flow across gramicidin-treated membranes is measured, corroborate this result. In 1 M salt solutions, streaming potentials are 2.35 mV per osmolal osmotic pressure difference instead of 3.0 mV. The smaller value may indicate multiple ion occupancy of the gramicidin channel at high salt concentrations. Apparent deviations from ideal cationic selectivity observed while attempting to measure single-salt dilution potentials across gramicidin-treated membranes result from streaming potential effects.


Water permeability of gramicidin A-treated lipid bilayer membranes. In membranes containing aqueous pores (channels), the osmotic water permeability coefficient, P f, is greater than the diffusive water permeability coefficient, P d. In fact, the magnitude of P f/P d is commonly used to determine pore radius. Although, for membranes studied to date, P f/P d monotonically declines with decreasing pore radius, there is controversy over the value it theoretically assumes when that radius is so small that water molecules cannot overtake one another within the channel (single-file transport). In one view it should equal 1, and in another view it should equal N, the number of water molecules in the pore. Gramicidin A forms, in lipid bilayer membranes, narrow aqueous channels through which single-file transport may occur. For these channels we find that P f/P d approximately 5. In contrast, for the wider nystatin and amphotericin B pores, P f/P d approximately 3. These findings offer experimental support for the view that P f/P d = N for single-file transport, and we therefore conclude that there are approximately five water molecules in a gramicidin A channel. A similar conclusion was reached independently from streaming potential data. Using single-channel conductance data, we calculate the water permeability of an individual gramicidin A channel. In the Appendix we report that there is a wide range of channel sizes and lifetimes in cholesterol-containing membranes.


Pre-lytic release of foot-and-mouth disease virus in cytoplasmic blebs. The pre-lytic release mechanism of foot-and-mouth disease virus was investigated by immunofluorescence, acridine orange staining, and electron microscopy in infected bovine and porcine kidney coverslip cultures. Cells with cytoplasmic fluorescence and which were positive for single stranded RNA with acridine orange staining were observed at 2 h after infection. Scanning electron microscopy showed cytoplasmic blebs in all cultures examined 2 h after infection. Rounded cells with virus inclusions began to appear 3 h after infection. Rounded cells and cytoplasmic blebs were shown to have single stranded RNA by acridine orange staining. Immunofluorescence and transmission electron microscopy with immunoferritin tagging demonstrated foot-and-mouth disease virus in cytoplasmic blebs. This study presents evidence for a pre-lytic release of foot-and-mouth disease virus through virus-containing cytoplasmic blebs emerging from infected cells.


Novel influenza A viruses isolated from Canadian feral ducks: including strains antigenically related to swine influenza (Hsw1N1) viruses. Twelve influenza A viruses, antigenically related to the Ho, H1 and Hsw1 subtypes, were isolated from cloacal samples of feral ducks in Canada. Antigenic comparisons showed that these viruses were most closely related to the recent HSW1N1 isolates from man and pigs, whereas in vivo pathogenicity tests revealed differences between the Hsw1N1 viruses from the ducks and those from humans and pigs. Antigenic characterization of 94 additional influenza A viruses from the ducks showed four haemagglutinin subtypes (Hav1, Hav4, Hav5 and Hav7), an unclassified haemagglutinin, and six neuraminidase subtypes (N1, N2, Neq2, Nav1, Nav2 and Nav5) in various combinations, some of which are novel and have not previously been reported. Three of these duck influenza viruses possessed a haemagglutinin antigenically related to that of classical fowl plaque virus. A much higher percentage of virus isolations were from juvenile ducks (18.5%) than from adults (5%). All of the ducks, from which viruses were isolated, appeared healthy at the time of sampling. Serological studies on a limited number of humans and domestic birds living in close proximity to the Canadian ducks revealed no evidence of interspecies transmission. Our findings suggest that these birds serve as a substantial reservoir of antigenically diverse influenza viruses, including isolates antigenically related to the current human and animal influenza viruses. This reservoir in nature may be perpetuated by a cycle involving annual infection of juvenile birds followed by transmission to the remaining susceptible birds until the next congregation during the breeding season.


Evidence for a precursor-product relationship between intracytoplasmic A particles and mouse mammary tumour virus cores. This report presents evidence which supports a relationship between intracytoplasmic A particles (CAP) and mouse mammary tumour virus (MMTV). Three MMTV-specific antigenic determinants in CAP (MMTV p27, p14 and p10) uere detected by immunodiffusion. No structural proteins of comparable mol. wt. were found in CAP; however, exposure of CAP to trypsin resulted in the cleavage of the CAP structural proteins to MMTV-like polypeptides. This process was accompanied by the preservation of MMTV-specific antigen determinants. Disulphide bonds were necessary for the structural maintenance of CAP. Reducing agents destroyed the organized structure of CAP, whereupon processing of CAP proteins to MMTV-like polypeptides by trypsin was prevented. CAP p82, possessed only MMTV p27 antigenic determinants, while CAP polypeptides p20--18 possessed p10 antigenic determinants. Following processing of CAP structural proteins by trypsin, MMTV-specific p27 antigenic determinants were shifted from CAP p82 to CAP p27; MMTV-p10 antigenic determinants were found with CAP p15--10. These results suggest a model wherein CAP structural proteins are modified by protease during maturation, resulting in the shift of their proteins to sizes consistent with those which have been currently identified as the major internal components of the virion and that this phenomenon is largely predicated on the folding of CAP proteins into the morphologically intact A particle.


Antigenic sites related to human serum proteins in HBsAg: isolation from normal human serum of a high-molecular-weight glycoprotein reacting with antialbumin. A novel minor constituent was isolated from normal human serum by affinity chromatography on columns of insolubilized concanavalin A and antibodies to albumin, followed by rate zonal centrifugation. This component has the following properties: a sedimentation coefficient of approximately 31; a diameter of 14--22 nm, and a buoyant density of 1.302 gm/cu cm. It contains about 1% neutral sugars and is electrophoretically heterogeneous, since two populations of particles with isoelectric points of pH 4.95 and 5.75 were separated by isoelectric focusing. It contains a single major glycopeptide with an apparent molecular weight of 72,000 daltons. Its amino acid composition is distinct from that of albumin. It elicited the formation of antibodies that also reacted with HBsAg.


Relationships between abnormal IgG index, oligoclonal bands, acute phase reactants and some clinical data in multiple sclerosis. The IgG-index and acute phase reactants were measured and oligoclonal bands were looked for in 30 patients with clinically definite multiple sclerosis (MS) and were compared with the clinical data. IgG-index was found elevated in 77% against 22% in a comparable material of patients with other neurological diseases. Oligoclonal bands were found in 74 and 17%, respectively. No correlation was found between these parameters and age, duration of illness, disability, coefficient of progression or age of onset. A statistical evaluation defines the specificity, sensitivity and validity of the methods used. CRP could no be demonstrated in 28, the rest had normal values. C3A was normal in all cases, but appeared to be elevated a little with increasing age, and to be positively correlated to the albumin-index, indicative of a defective blood-brain barrier. No correlation was found between CRP, C3A and the clinical data.


Giant cell arteritis (cranial arteritis, polymyalgia rheumatica). Giant cell arteritis, which is probably due to disturbed immune mechanisms, has a spectrum of clinical symptoms in elderly people. In nearly all cases such general signs as loss of appetite, loss of weight and fever are present. The sedimentation rate is almost without exception about 100 mm in the first hour. The two most frequent and typical clinical syndromes are polymyalgia rheumatica and cranial arteritis. The polymyalgia rheumatica is characterized by periarticular pain which is mostly symmetrical and accentuated in the shoulder girdle. Increasingly severe temporal headache and ocular distrubances are found with cranial arteritis in more than 50% of cases. A combination of both diseases is frequent. Other arterial branches are rarely involved. The course of the disease is over a period of 1 1/2 to 2 years. Treatment with corticosteroids is indicated mainly because of the severe ocular complications with blindness. It should begin immediately, be intensive and last over a long period. Regular followup is necessary over several years in order to avoid relapses.


The enhancing effect of multiple sclerosis brain homogenates on the active E rosette forming lymphocytes. Lymphocytes from 15 of 17 patients with clinically well defined multiple sclerosis (MS) responded to all four MS brain homogenates as assayed by active E rosette test. Lymphocytes from only one neurological patient, out of 18 tested, responded to MS brain homogenates. The nature of the stimulating agent from MS brains remains to be determined.


Transient muscular weakness in severe recessive myotonia congenita. Improvement of isometric muscle force by drugs relieving myotomic stiffness. The maximum force of voluntary muscle contraction was registered under isometric conditions in nine patients with recessive myotonia congenita. The recordings were made on the upper arm. Five patients with severe myotonia had a transient weakness after muscle rest. Electromyographic registrations with wire electrodes showed that the myotonic muscle fiber discharges disappeared during the transient weakness. Medication improving myotonic stiffness also improved the weakness. The cause of transient weakness seems to be similar to that of myotonic stiffness. It is known that an increasing depolarization of the myotonic muscle fiber membrane leads to the myotonic discharges and myotonic stiffness. In severe myotonia the progressing depolarization could cause a loss of excitability of the muscle fiber membrane and thereby a transient paresis of a more or less large number of muscle fibers.


Hereditary bimanual synkinesis combined with hypogonadotropic hypogonadism and anosmia in four brothers. A new genetic syndrome of the combined occurrence of hypogonadotropic hypogonadism, anosmia (Kallmann syndrome) and congenital mirror movements in four brothers is presented. Mirror movements were manifest only within the distal parts of the upper extremities and resembled congenital mirror movements described for isolated or familial cases or those occurring in combination with other genetic defects. The hypothesis is supported, that a midline fusion disorder with preponderance of uncrossed pyramidal tract fibers is a major pathogenetic factor for the occurrence of congenital mirror movements.


Microsphere analysis of local cerebral and extracerebral blood flow after complete ischemia of the cat brain for one hour. In normothermic anesthetized cats complete cerebral circulatory arrest for one h was produced and followed by blood recirculation of the brain for 30 min to 4 h. Total and local blood flow of the brain, kidney, heart and liver were measured before and after ischemia using radioactive labelled microspheres. Before ischemia blood flow of the brain was 39.1 +/- 2.3 ml/100 g/min, of the kidney 307.2 +/- 28.3 ml/100 g/min, of the heart 241.1 +/- 32.5 ml/100 g/mmin and of the liver 87.8 +/- 25.6 ml/100 g/min (means +/- SEM). Regional flow rates within the brain varied between 35 and 51 ml/100 g/min. Reactive hyperemia was present in the brain 30 min after the beginning of recirculation following ischemia for 1 h. Local cerebral flow rates increased three to five times above the control flow, depending on the respective region. Mean cerebral blood flow returned to or slightly below normal, 2 to 4 h after ischemia, but there was considerable redistribution of flow rates within the brain. The filter capacity of the brain for microspheres of 15 mu and 50 mu diameter did not change after ischemia indicating that postischemic blood recirculation was not accompanied by an opening of arteriovenous shunts.


Retrograde changes of nerve fibers with the carpal tunnel syndrome. An electroneurographic investigation. In the CTS the motor nerve conduction velocity proximal to the wrist is reduced in proportion to the degree of severity of the nerve lesion. Furthermore the evoked nerve action potential is significantly reduced when recordings are made from the median nerve at the elbow and when the compound nerve is stimulated proximal to the lesion at the wrist. The extent of the retrograde changes correlates with the degree of severity and duration of nerve compression. Measurement of the evoked nerve action potential in the proximal nerve segment enables us to estimate the extent of the retrograde nerve fiber degeneration and therefore might be important for prognosis.


Encephalomyelitis with thyrotoxicosis.

Encephalomyelitis with thyrotoxicosis. A case of thyrotoxicosis associated with neuropathy and encephalomyelitis is reported which gradually improved as regards the hyperthyroidism and the neurological deficit during treatment with Tapazole. The possible role of the nervous system of an excess of thyroxine or an autoimmune factor as a cause of the involvement is discussed.


Interaction of nitrofurantoin with diphenylhydantoin. Left-sided motor seizures in a patient with an operated brain tumor were controlled with 300 mg/d DPH. The introduction of antimicrobial therapy with nitrofurantoin caused a fall of serum DPH levels and the recurrence of seizures, at the same time serum gammaGT values were increased. These changes were reversible after nitrofurantoin treatment was terminated. Both increased DPH metabolism or impaired absorption could be responsible for this effect. The concomittant increase of gammaGT could be interpreted as indirect evidence of hepatic enzyme induction with increased metabolism of DPH. It is important to note the possibility of such interaction so that better anticonvulsant control can be achieved.


Consciousness and disturbances of consciousness. The problem of consciousness is discussed briefly, including the contrary views of consciousness as a transcendental phenomenon and as an animistic fiction. Measurement of consciousness is possible only indirectly by means of quantitative assessment of accompanying behavioral deficits. Knowledge of the structural basis of consciousness is incomplete. The ascending reticular activating system (ARAS) is necessary for the maintenance of the state of consciousness. The monoamine and a great number of descending projections modulate the ARAS. The contents of consciousness depend also on telencephalic structures, primarily on the telencephalic cortex. Certain localized telencephalic lesions bring about disturbances of consciousness. The role of the corpus callosum in the problem of consciousness is discussed (one brain--two minds hypothesis). Then a classification of the various disturbances of consciousness is proposed. The term "disturbances of vigilance" is used for all disturbances of consciousness which are caused by a lesion in or a functional disorder of the ARAS or any of its modulating subsystems. The term "disturbances of the contents of consciousness" refers to disturbances of consciousness due to global or localized lesions or functional disorders of telencephalic structures. A list of characteristic features is given for each class of disturbances of consciousness.


Clinical studies of the movement-related cortical potential (MP) and the relationship between the dentatorubrothalamic pathway and readiness potential (RP). In order to investigate the influence of basal ganglia and cerebellar involvement on the preparatory state of the cerebral cortex for voluntary movement, the cortical potential preceding finger movement was studied in 20 patients with Parkinson's disease and 20 patients with cerebellar ataxia. Readiness potential (RP) was abnormal in 90% of the Parkinson group and in 55% of the cerebellar ataxia group. The most frequent abnormality was a depressed amplitude and earlier onset of RP in both groups. The most remarkable finding in the present study was the complete absence of RP with dyssynergia cerebellaris myoclonica (presumed Ramsay Hunt syndrome) whereas normal RP was obtained with cerebellar cortical degeneration. In addition, RP was absent or severely depressed in patients with a unilateral vascular lesion of the midbrain (Benedikt's syndrome) and in patients with Parkinson's disease who underwent unilateral intermedioventral (Vim) thalamotomy. These facts suggest a possible important role of the dentatorubrothalamic or dentatothalamic pathway in the physiogenesis of RP.


The geographical distribution of multiple sclerosis, rheumatoid arthritis, rheumatic heart disease and poststreptococcal nephritis in Sardinia: climatic and socioeconomic factors. The comparative geography of multiple sclerosis (MS) and nonneurological diseases considered to be autoimmune is of great interest. But there are few appropriate investigations. Some have found an increase in the frequency of MS, rheumatoid arthritis and rheumatic heart disease depending on the geographic latitude. Nevertheless, the significance of the latitude effect as an indicator of a possible etiological relationship between MS and these conditions has been questioned. In this paper, the frequency of MS, rheumatoid arthritis, rheumatic heart disease and poststreptococcal nephritis is reported from Sardinia where appreciable differences in climatic and socioeconomic conditions exist. There was a positive correlation of the distribution of MS with the distribution of rheumatic heart disease and poststreptococcal nephritis. On the other hand, no correlation was found with the distribution of rheumatoid arthritis.


Nemaline bodies in a progressive infantile neuromuscular disorder. Nemaline bodies are described in a case of a progressive neuromuscular disorder, which is difficult to classify. The clinical syndrome is not characteristic of a nemaline myopathy. It is argued that the finding of nemaline bodies is in itself not specific of any neuromuscular disease.


Thalamic EEG recordings in patients with chronic pain. Thalamic EEG recordings were made in 10 patients; the therapeutic goal of the implantation was the electrical stimulation treatment for pain. The patients' ages ranged from 37 to 72 years; seven patients had thalamic pain (Dejerine-Roussy syndrome), two had chronic spinal arachnoiditis and one had facial anaesthesia dolorosa. Platinum-iridium electrodes were stereotaxically inserted; the ventrobasal complex of the thalamus including the ventral posterior zone was the target of the implant. Excessive thalamic slowing was found in four of seven patients with Dejerine-Roussy syndrome and also in two of three cases with other causes of pain. One patient had marked rhythmical intermittent delta activity in the thalamus which was often triggered by arousing stimuli. Thalamic spindle activity was sometimes noted without concomitant spindle activity on the scalp and would occasionally occur in states of early drowsiness.


Hyperreactive (hyaline, opaque, dark) muscle fibers in Duchenne dystrophy. A biopsy study of 16 dystrophy and 205 other neuronmuscular disease cases and controls. Biopsies of 16 patients with Duchenne dystrophy and 205 consecutive biopsies of other neuromuscular diseases and controls were studied for the presence of hyperreactive (hyaline, dark, opaque) fibers. All biopsies of Duchenne dystrophy showed hyperreactive fibers; the percentage varies between values below 1 up to 19 (median 3.5%). In 11 (69%) of the 16 biopsies the percentage was more than 1. Only in 34 (17%) of the 205 consecutive biopsies were hyperreactive fibers found. In 30 of these 34 biopsies the percentage of hyperreactive fibers was below 1. A positive or negative correlation between the increased level of serum-CPK or the early stage of Duchenne dystrophy and the number of hyperreactive fibers could not be demonstrated. Obviously the hyperreactive fibers are not specific for Duchenne dystrophy. On the other hand a value of 1% supports this diagnosis. Delta lesions were demonstrated in 54% of the hyperreactive fibers.


Evaluation of the jitter phenomenon in myasthenic patients and their relatives. Single fiber electromyography with jitter measurements and blocking evaluation was used for investigating 23 myasthenic patients, their 29 healthy relatives, coming from 10 families, and a control group of 10 subjects. Blocking was never seen in the healthy controls, and the jitter averaged some 30 microsec. Blocking was present in the myasthenic patients in 25.4% of the recorded potential pairs and there was a pathological jitter, which often extended over a few hundred microsec. The records of ten healthy relatives of patients were suggestive of slight but unquestionable abnormalities of neuromuscular transmission, viz. blocking and a jitter of over 60 microsec or more. The results demonstrate subclinical derangements of neuromuscular transmission in families of myasthenic patients, which points to a familial factor as involved in the pathomechanism of the disease.


Antifibrinolytic therapy of subarachnoid hemorrhage by intrathecal administration of p-aminomethylbenzoic acid. In patients suffering from subarachnoid hemorrhage (SAH), intrathecal administration of p-aminomethylbenzoic acid (PAMBA) was found to produce hemostatically effective concentrations of the antifibrinolytic agent in the cerebrospinal fluid (CSF). The early clinical findings speak in favor of this mode of administration.


Myotonia induced with clofibrate in rats. Myotonia was induced in rats with clofibrate given in daily subcutaneous injections of 0.4 g/kg. The first myotonic discharges were recorded electromyographically from the extensor digitorum longus, tibialis anterior, and gastrocnemius muscles after 4 days on clofibrate, but from the soleus not until after 11 days. Clofibrate induced myotonic activity in chronically denervated muscle also. During repetitive nerve stimulation the electrical response of the muscle was declining in all myotonic rats. It did so also when repetitive stimulation was applied directly to the muscle, which would seem to suggest a myotonic defect as the cause. Several drugs were tested and diphenylhydantoin proved to inhibit myotonia most effectively. Animals on an extended clofibrate schedule (12 weeks) had ECG abnormalities resembling those seen in patients with myotonic dystrophy.


Anterior fusion for cervical spondylosis. Anterior fusion was performed on 138 patients as treatment for degenerative changes of the cervical vertebral column. The results were checked after up to 11 years in 122 patients and were found to be good in 55%, fair in 3895% and poor in 6.5%. There were complications of phonetic paralysis in one case, Horner syndrome in two cases, 2 wound infections, and complaints about the iliac crest in 3 patients. Kyphosis at the fused segment occurred 26.1% of cases, the average angle being 15.3 degrees, but it did not influence the clinical results. Mortality was 2%. Inadequate visualization of the nerve roots at operation was probably the reason for the segmental deficits and suggestions were made to avoid this by EMG, microsurgical technic, uncusectomy and hemifacetectomy. The results in the presence of myelopathy were much poorer, presumably because of associated degenerative changes in the older patients.


Screening of patients with Huntington's chorea by pneumoencephalography: criteria for decision and probability of error. The problem of making a decision in case of two clinical hypotheses is treated by the application of the Neyman-Pearson criterion. As an example, a pneumoencephalographic index (frontal horn width divided by septum-caudate distance) of two different diseases, Huntington's chorea and Parkinsonism, is analyzed. As the aim of the investigator is to detect the maximum number of cases of Huntington's chorea, an appropriately high alpha, (the probability of "false alarm") should be chosen, and maximization of 1-beta (the probability of detection of Huntington's chorea) carried out.


Myotonic myopathy with painful muscle contractions and decrease of symptoms by cold. Myotonic reaction and paresis accompanied by painful muscle contractions limited to the upper limbs, which decrease remarkably in the cold, were observed in a 29 year old man. The histological investigation revealed minimal non-specific signs of myopathy. The biochemical studies of muscular tissue contained a normal amount of myophosphorylase, acid maltase and glycogen. Ischemic work induced normal elevation of venous lactate. The activities of CPK, LDH and SGOT in the blood serum were occasionally increased. The EMG showed typical myotonic bursts and electrical silence during painful muscle contractions. Repetitive high frequency stimulation demonstrated a clear initial increase of the amplitude of action potentials followed by a decrease in the contracted muscle. The father of the patient suffered from dystrophia myotonica. This coincidnece suggests that this myotonic myopathy is a variant of dystrophia myotonica.


Oligodendroglioma with a twenty-two year history. Clinicopathological case report. Clinico-pathologic report of a patient with an oligodendroglioma who ran a 22-year course characterized by focal epileptic seizures of the Jaksonian type. The EEG remained normal for many years. Since the tumor had not been influenced by therapeutic measures, the histologic picture permitted considering some basic problems of origin, growth, and differentiation of oligodendrogliomas.


A case of epidermoid of the fourth ventricle. The case of woman, aged 52, who had a fourth ventricular epidermoid, which eluded definite diagnosis for six years because of spontaneous remissions, is reported. Computed tomography may be misleading in such cases with spontaneous remission of symptoms because of the low electron density of the tumor which is not enhanced by intravenous contrast medium. Other diagnostic means such as ventriculography should not be neglected.


alpha-Albumin (glial fibrillary acidic protein) in normal and pathological human brain and cerebrospinal fluid. alpha-Albumin, a specific brain protein observed after agar gel electrophoresis and shown to be identical to the later described GFA, has been determined in normal and pathological human central nervous system and cerebrospinal fluid. The outcome of this study underlines the value of the detection of specific proteins of the brain in biological fluids.


Eosinophilic granulocytes in cerebrospinal fluid: analysis of 94 cerebrospinal fluid specimens and review of the literature. Eosinophilic granulocytes in the CSF were observed in 94 of approximately 10,000 qualitative cytologic preparations. Those cases of eosinophilia which occurred in the context of a parasitic disease or a puncture-related hemorrhage were excluded. CSF eosinophilia exceeding 1% was found in 57.5% of the cases and 5% in 23.5%. Increased cell counts were observed in 67.7% of the cases; elevated CSF protein values, in 68% to 73%; blood eosinophils, in 10.4%. There was no reason to suspect a relationship between these findings and the number of eosinophils in the CSF.--Fifty-two percent of the cases involved inflammatory diseases of the nervous system; the 18 cases of abacterial inflammation of unknown etiology were particularly striking. In the remaining cases, eosinophils were found in conjunction with cerebral ischemia and hemorrhage, with tumors, and in a relatively high percentage of children (21%). The frequency of occurrence with drained or undrained hydrocephalus was striking. A review of the pathophysiological function of eosinophils indicated that revived or corpuscular antigens were present in all cases of CSF eosinophilia in which an eosinophilic reaction was induced. Nothing can be said at this time, however, concerning the classification of the antigens.


Hepatic synthesis and urinary excretion of alpha2u-globulin by male rats: diurnal rhythm and response to fasting and refeeding. The urine of sexually mature male rats contains a protein of hepatic origin, alpha2u-globulin, not found in the urine of immature or female rats; output of this protein is greatly reduced by fasting. We have examined the effects of feeding and of fasting for various lengths of time on urinary output and hepatic synthesis of alpha2u-globulin. Rats eating ad libitum showed diurnal rhythms of urinary alpha2u-globulin excretion reaching maxima between 2000 and 0800 hours, thus coinciding with the daily feeding period of the rat. Fasting for 12 hours extinguished this diurnal rhythm. When fasting was prolonged up to 36 hours, urinary excretion of alpha2u-globulin was reduced to very low levels. Hepatic synthesis of alpha2u-globulin under these nutritional conditions was investigated by incubating liver polyribosomes with [3H]leucine and a preparation of soluble enzymes for protein synthesis and separating the labeled alpha2u-globulin peptides by immunoprecipitation followed by electrophoresis on sodium dodecyl sulfate-polyacrylamide gels. By this technique, it was shown that only membrane-bound ribosomes in the livers of mature male rats make this protein. Semi-quantitative measurement suggested that the proportion of liver polyribosomes synthesizing alpha2u-globulin was unchanged after 12 hours of fasting, but was reduced after 24 and 36 hours of fasting. It is proposed that the diurnal rhythm in alpha2u-globulin output in the urine represents translational control of its synthesis in the liver, whereas the more extensive reduction with prolonged fasting is partly due to a selective reduction in transcription of the messenger RNA for this protein.


The longitudinal course of congenital rubella encephalitis in nonretarded children. The neurologic course of congenital rubella syndrome was traced in 29 nonretarded children to 9 to 12 years. During the first two years, manifestations involved abnormal tone and reflexes (69%), motor delays (66%), feeding difficulties (48%), and abnormal clinical behavior (45%). Hearing loss was documented in 76%. From three to seven years, poor balance, motor incoordination (69%), and behavioral disturbances (66%) predominated. Hearing losses increased to 86%. Currently, at 9 to 12 years, 25 have residua which include learning deficits (52%), behavioral disturbances (48%), poor balance (61%), muscle weakness (54%), and deficits in tactile perception (41%). Two additional children now have hearing loss. The encephalitic manifestations of congenital rubella syndrome are diverse. Overriding problems differ at each phase of childhood. Current deficits influence progress in educational and home environments. For these children, adequate intelligence alone does not guarantee academic success.


Fast axoplasmic transport of noradrenaline and dopamine in mammalian peripheral nerve. A fast transport of noradrenaline (NA) at a velocity of 392 mm/day was found in cat peroneal nerve using a double-ligation technique and a new sensitive radioenzymatic assay for catecholamines. The velocity of transport of NA is sufficiently close to that of 410 mm/day found for labelled proteins and polypeptides to be considered as moving down within the nerve fibres by the same transport mechanism. In addition, dopamine (DA) was also found to be moved down by anterograde transport at a fast, but not well defined, rate. Disulfiram, a dopamine-beta-hydroxylase inhibiting agent, reduced NA levels and increased DA both in control nerve segments and within the portion of the nerve isolated by ligations where NA-containing densecore vesicles are present. The results are considered to support a dynamic turnover of NA and DA in the dense-core vesicles as they are trasnported in the axons.


Kinetic properties of normal and perturbed axonal transport of serotonin in a single identified axon. 1. The axonal transport of pulses of [3H]serotonin was studied in an axon of the serotonergic giant cerebral neurone (GCN) of Aplysia californica. 2. [3H]serotonin was transported as a discrete peak which was followed by a relatively low, smooth trail. 3. The peak broadened as it moved along the axon, sometimes skewing in the proximal direction. 4. The velocity of the transport was highly dependent on temperature, but the rate of peak broadening was not. The velocity was 130 mm per day at 23 degrees C and 48 mm per day at 14 degrees C. The rate of broadening was 143 micrometer per mm transport at 23 degrees C and 156 micrometer per mm transport at 14 degrees C. 5. In another series of experiments, almost the entire length of the lip nerve, which contained the axon of GCN, was maintained at 1--3 degrees C to block transport. The GCN's cell body and the proximal few millimetres of the nerve were maintained at 23 degrees C. As a result, the amount of [3H]serotonin in the proximal segment of the nerve increased manyfold during periods of up to 4 hr. The concentrated pulse of [3H]serotonin resulting from this treatment was transported more slowly than normal after the cooling was terminated. Sometimes, a minor peak split from the major peak of radioactivity and was transported a normal velocity. 6. Incubation of the cerebral ganglion and nerves for 16 hr in the presence of anisomycin, an inhibitor of protein synthesis, reduced by nearly fourfold the amount of [3H]serotonin subsequently exported into the axon of the GCN. The transport velocity at this reduced concentration was less than half the normal value. If the concentration of [3H]serotonin in the axon was restored to normal in the presence of anisomycin, the velocity of transport was also returned to normal. 7. We conclude that the velocity of transport of serotonergic vesicles in the axon of the GCN is positively dependent on the local concentration of vesicles, except at very high concentrations, where the dependence is negative. The results are interpreted in the context of a model for transport in which the serotonergic vesicle is translocated along the axon in an intermittent fashion, alternating between moving and stationary states. The local concentration of the vesicles along the axon would control the observed velocity of transport by altering the partitioning between the two states, that is, by changing the percentage of time vesicles spend in each state.


Influence of illness on developmental progress in children under two. Illnesses occurring in children under two were recorded in relation to a systematic programme of child surveillance in one general practice.The most important result of this study was that although illness does have a detectable effect on developmental progress, the degree of that influence is for the most part small.Illnesses experienced between 0 and 7 months have less influence upon seven-month developmental scores than upon 24-month developmental scores, but have a greater influence upon the latter than do illnesses experienced between 7 and 24 months.There was a significantly higher consultation rate for boys and for children from social classes 3b, 4, and 5.


Chondroma rodens in a dog.

Chondroma rodens in a dog. The clinical and pathological characteristics of a chondroma rodens affecting the internal occipital protruberance, cerebellar meninges and the cerebellum of a dog, are described. The principle clinical findings were Horner's syndrome and ataxia. The histopathologic differentiation of this tumour from other fibro-osseous growths, is discussed.


Effect of calcium on the H+/K+ ATPase of hog gastric microsomes. The K+-stimulated, ouabain-insensitive ATPase activity present in vesicles of microsomal fractions from hog gastric mucosa can be demonstrated in fresh preparations by adding Ca2+ (micron range) to the incubation medium. Ca2+ effect is similar but not additive to the effect of gramicidin or freezing. High Ca2+ concentrations (1 mM) produce an inhibitory effect on the K+-stimulated ATPase activity. This effect is not seen in the presence of gramicidin. Calcium increases the magnitude of ATP-driven H+ uptake in vesicles exposed to K+ for periods of time up to 60 min. At longer times of exposure (120 min) the response does not differ from controls. It is concluded that Ca2+ at low concentrations (micron range) enhances the K+ permeability of the vesicular membrane. At higher concentrations (mM range), Ca2+ becomes inhibitory to the K+ permeability. A role for Ca2+ as a second messenger in stimulus-secretion coupling in the parietal cell is discussed.


Purification and characterization of a human lung tumor-associated antigen. A human lung tumor-associated antigen was purified from a saline extract of a lung adenocarcinoma. The antigen was demonstrated in extracts of lung adenocarcinoma. The antigen was demonstrated in extracts of lung tumors with the use of an absorbed antiserum by double-diffusion immunoprecipitation. The antiserum did not react with extracts of normal lung or other normal tissues, and the antigen was immunologically distinct from other tumor-associated antigens. Purification was achieved by antibody affinity chromatography and preparative polyacrylamide gel electrophoresis. Isolation procedures were monitored by immunoreactivity with absorbed monospecific antiserum. The antigen was labeled with 125I and judged homogeneous by 1) polyacrylamide gel elecrophoresis in detergent and nondetergent gels, 2) molecular sieve chromatography, 3) ion exchange chromatography, and 4) sucrose gradient sedimentation analysis. A molecular weight of 77,000 was calculated from the s20.w value of 4.24S and from the D20.w value of 5.0X10(-7) cm2/sec. Sodium dodecyl sulfate gel electrophoresis indicated a subunit molecular weight of 42,000. The Stokes radius of the antigen was 40 A and the frictional ratio was 1.42, indicating a nonspherical molecule. The purified radioiodinated antigen could be quantitatively precipitated with specific antiserum.


Rabbit antiserum against a non-T, non-B leukemia cell line that carries the Ph1 chromosome (NALM-1): antibody specific to a non-T, non-B acute lymphoblastic leukemia antigen. Rabbit immune sera against human cells of a non-T, non-B leukemia cell line bearing the Ph1 chromosome (NALM-1) were characterized. After proper and exhaustive absorption, the sera were tested by indirect membrane immunofluorescence on a panel of 19 human hematopoietic cell lines with T-, B-, or non-T, non-B cell surface characteristics. The absorbed sera reacted specifically with NALM-1 cells but not with cells of another Ph1-positive cell line, K-562. Four of the 6 leukemia T-cell lines, 2 of the 4 leukemia-lymphoma B-cell lines, all of 4 acute lymphoblastic leukemia (ALL) lines with non-T, non-B cell characteristics, and uncultured cells of all patients with non-T, non-B cell ALL were reactive with the antisera. A cross-absorption study of the antisera suggested that a single antigenic complex is involved in this antigen specificity. The antigen involved appears to be a common non-T, non-B ALL one that has been described previously.


The surgical management of patients with carcinoma of the midthoracic esophagus. Clinical and pathological findings in 15 patients with carcinoma of the midthoracic esophagus were analyzed. The author concluded that preoperative radiotherapy reduces tumor size and improves resectability. Longer follow-up is required to determine whether or not preoperative radiotherapy improves survival rates in these patients.


Abate--its value as a cyclopscide.

Abate--its value as a cyclopscide. Guinea worm disease is highly endemic in Kurnool district in Andhra Pradesh. The effect of Abate on the vector cyclops was studied and was found to be very encouraging.


Inhibition by RNA of RNase H activity associated with reverse transcriptase in Rauscher murine leukemia virus cores. We reported earlier that core preparations of Rauscher murine leukemia virus, when separated on an isopycnic sucrose gradient, did not contain detectable levels of RNase H activity, while retaining high levels of reverse transcriptase activity. We reexamined this phenomenon, and the earlier observation was found to be reproducible. However, when doubly banded preparations of viral cores were solubilized and reverse transcriptase was isolated by ion-exchange chromatography, a coincident peak of a nuclease activity with the specificity of RNase H was observed, which indicated that RNase H was selectively inhibited in the core fractions. By direct activity measurements using the purified reverse transcriptase-RNase H from cores, this endogenous inhibitor has been identified as the viral RNA. Viral 70S RNA strongly inhibited RNase H activity purified either from whole virions or from prefractionated cores. Other RNAs tested that had inhibitory effects were yeast tRNA, polyadenylic acid, and polyguanylic acid. Polyuridylic acid and polyadenylic acid were moderately inhibitory, and polycytidylic acid did not inhibit the RNase H. A rabbit anti-reverse transcriptase immunoglobulin G inhibited both the reverse transcriptase and RNase H activities of the enzyme purified from cores. These data provide a rational explanation for the failure to detect RNase H activity in core preparations of Rauscher murine leukemia virus. Furthermore, these data are consistent with the idea that the RNase H and reverse transcriptase activities purified from cores reside on the same protein molecule. Possible biological implications of the observed inhibition of RNase H by RNA is discussed.


Reverse transcriptase-associated RNase H activity. II. Inhibition by natural and synthetic RNA. The RNase H activity associated with purified avian myeloblastosis virus and Rauscher murine leukemia virus DNA polymerases is inhibited by homopolymeric RNA molecules, although the efficiency of inhibition by each homopolymer appears enzyme specific. Formation of duplex RNA molecules abolished the inhibitory activity. In contrast to these results, DNA polymerase-independent RNase H activities, such as the RNase H-II from Rauscher murine leukemia virus and calf thymus RNase H, were unaffected by the addition of exogenous RNA molecules to reaction mixtures. These results support the concept (M. J. Modak and S. L. Marcus, J. Virol. 22:253--256, 1977) that the catalytic site of DNA polymerase-associated RNase H activity may be that which is also involved in template binding. Naturally occurring RNA molecules of oncornaviral, procaryotic, or eucaryotic origin also proved to be efficient inhibitors of avian myeloblastosis virus DNA polymerase-associated RNase H activity. In contrast to this result, naturally occurring RNA molecules, at concentrations which inhibited the avian myeloblastosis virus enzyme, did not inhibit Rauscher murine leukemia virus DNA polymerase-catalyzed RNase H activity. This finding represents a new biochemical distinction between the two reverse transcriptases, and may be indicative of differences in the relative affinities of these enzymes for natural RNA molecules.


Mammalian retrovirus-associated RNase H is virus coded. RNase H of a temperature-sensitive mutant of Rauscher murine leukemia virus is thermolabile, establishing this activity as a virus-coded function of the mammalian type C virus reverse transcriptase.


Analysis of translational products of Friend strain of spleen focus-forming virus. We have analyzed normal rat kidney cells nonproductively infected with the Friend strain of spleen focus-forming virus (SFFV) for the presence of murine leukemia virus-specific type C viral proteins. SFFV was found to code for the p15 and p12 proteins of Friend-murine leukemia virus as determined by immunological typing of their antigenic determinants. Molecular-size analysis of p15 and p12 proteins in SFFV nonproductively infected normal rat kidney cells indicated that these proteins are translated as parts of polyprotein molecules. The apparent molecular weights of the polypeptides bearing p12 antigenic determinants revealed the presence of translational products of the SFFV genome that could not be accounted for by know type C virus helper structural proteins.


Virus-coded origin of a 32,000-dalton protein from avian retrovirus cores: structural relatedness of p32 and the beta polypeptide of the avian retrovirus DNA polymerase. A 32,000-dalton protein (p32) located in avian retrovirus cores was immunoprecipitated from [35S]methionine-labeled avian myeloblastosis virus (AMV) propagated in cultured chicken embryo fibroblast cells by an antiserum preparation (sarc III) derived from tumor-bearing hamsters injected with cloned and passaged cells from an avian sarcoma virus-induced primary hamster tumor. Since sarc III serum apparently contained antibodies only to virus-coded proteins and not to chicken cellular proteins, the immunoprecipitation of p32 from AMV by sarc III serum strongly suggested that p32 is virus coded. The origin of p32 was more definitively established by demonstrating the existence of a structural relationship between p32 and the AMV DNA polymerase. AMV p32 cross-reacted with the beta polypeptide of AMV alphabeta DNA polymerase in radioimmunoprecipitation and radioimmunoprecipitation inhibition assays, indicating that p32 and beta share common antigenic determinants. This relationship was clarified by sodium do-decyl sulfate-polyacrylamide gel electrophoretic analysis of the peptides generated by limited proteolysis of 125I-labeled AMV DNA polymerase polypeptides and of 125I-labeled AMV p32 by chymotrypsin or Staphylococcus aureus V-8 protease. The peptides which appeared during proteolytic digestion of p32 were a subset of those produced by digestion of the beta polypeptide; however, p32 had no discernible peptides in common with the alpha polypeptide. Further, all of the peptides produced by limited proteolysis of beta were present in the digests of either p32 or alpha. Our findings suggest that p32 is apparently derived by cleavage of the beta polypeptide of AMV DNA polymerase, presumably at a site near or identical to that at which alpha is generated from beta by proteolytic cleavage.


Molecular diversity among five different endogenous primate retroviruses. Genetically transmitted retroviruses of Old and New World monkeys include type C viruses isolated from baboons (M7), macaque (MAC-1), and owl monkeys (OMC-1) and type D viruses from langurs (PO-1-Lu) and squirrel monkeys (SMRV, M534). Each of these isolates is unrelated to the others by nucleic acid hybridization criteria and contains a unique array of virion-associated proteins which can be resolved by agarose gel filtration and polyacrylamide gel electrophoresis under denaturing conditions. The major structural protein of each virus has a distinct primary structure, as determined by two-dimensional tryptic peptide analysis, and is antigenically different from the others. The major virion phosphoproteins of endogenous primate type C viruses (pp15) are also different from those of type D viruses (pp13-pp14). Immunological and structural analyses show that the endogenous langur virus and the horizontally transmitted Mason-Pfizer virus of rhesus monkeys are closely related to one another, consistent with the sequence homology detected in their RNA genomes. Although certain radioimmunoassays detect interspecies antigenic determinants common to either the p30 or gp70 proteins of some of these viruses, no one assay has yet been designed which can detect all groups of endogenous primate retroviridae. The data lead to the conclusion that primates contain a minimum of three different sets of genetically transmitted type C and type D retroviral genes.


Characterization of K virus and its comparison with polyoma virus. The antigenic relationship between the two murine papovaviruses, K virus and polyoma virus, was examined by serological techniques to determine whether they shared any antigenic components. No cross-reactivity was found associated with the viral (V) antigens by the indirect immunofluorescence, neutralization, or hemagglutination-inhibition tests. The tumor (T) antigens expressed in transformed cells or cells productively infected by either K or polyoma virus did not cross-react by indirect immunofluorescence. An antigenic relationship was detected, however, among the late proteins of K virus, polyoma virus, simian virus 40, and the human papovavirus BKV, when tested with either hyperimmune sera prepared against polyoma virus and simian virus 40 or sera prepared against disrupted virions. The nucleic acids of K and polyoma viruses were compared by agarose gel electrophoresis and restriction endonuclease analysis. No nucleotide sequence homology between the genomes of these two viruses was detectable by DNA-DNA hybridization techniques under stringent conditions. The genome of K virus was found to be slightly smaller than that of polyoma virus, and the cleavage patterns of the viral DNAs with six restriction endonucleases were different. These findings indicate that there is little relationship between these two murine papovaviruses.


Isolation and characterization of a virus-specific ribonucleoprotein complex from reticuloendotheliosis virus-transformed chicken bone marrow cells. Chicken bone marrow cells transformed by reticuloendotheliosis virus (REV) produce in the cytoplasm a ribonucleoprotein (RNP) complex which has a sedimentation value of approximately 80 to 100S and a density of 1.23 g/cm3. This RNP complex is not derived from the mature virion. An endogenous RNA-directed DNA polymerase activity is associated with the RNP complex. The enzyme activity was completely neutralized by anti-REV DNA polymerase antibody but not by anti-avian myeloblastosis virus DNA polymerase antibody. The DNA product from the endogenous RNA-directed DNA polymerase reaction of the RNP complex hybridized to REV RNA but not to avian leukosis virus RNA. The RNA extracted from the RNP hybridized only to REV-specific complementary DNA synthesized from an endogenous DNA polymerase reaction of purified REV. The size of the RNA in the RNP is 30 to 35S, which represents the subunit size of the genomic RNA. No 60S mature genomic RNA was found within the RNP complex. The significance of finding the endogenous DNA polymerase activity in the viral RNP in infected cells and the maturation process of 60S virion RNA of REV are discussed.


Papain solubilization of the Epstein-Barr virus-induced membrane antigen. The Epstein-Barr virus (EBV)-induced membrane antigen (MA) was successfully solubilized from the membranes of viable EBV-infected Raji cells by treatment with papain (5 to 6 U per 1 X 10(7) to 2 X 10(7) cells). The loss of MA from viable cells was monitored by membrane immunofluorescence and antibody-dependent cellular cytotoxicity. Soluble MA was demonstrated in papain digests through inhibition of antibody-dependent cellular cytotoxicity and by inhibition of the binding of anti-MA antibodies to cells as detected by use of 125I-labeled staphylococcal protein A. Approximately 75% of the MA activity in the extracts was not sedimentable at 100,000 X g,, indicating that the majority of EBV MA activity that was released by this procedure was associated with small-molecular-weight material. Antiserum prepared from an owl monkey immunized with these papain extracts contained antibody to MA and neutralizing antibodies, but lacked detectable antibodies against viral capsid antigens and EBV-induced early antigens.


A comparison of serum immunoglobulins from patients with non-neoplastic prostates and prostatic carcinoma. The major immunoglobulin classes were surveyed among 23 patients with carcinoma of the prostate, 14 patients with clinically manifest benign prostatic hyperplasia and 23 healthy, elderly men without evidence of prostatic disease to determine if differences in immunoglobulin levels existed. Levels of IgG,IgA and IgM were determined by single radial immunodiffusion. Serum IgM levels were depressed in patients with all stages of carcinoma of the prostate as compared to levels in controls. These depressions were significant statistically for the tumor group considered as a whole and for patients with stages A and B tumors; the depression of IgM levels in patients with stages C and D tumors bordered on statistical significance. Serum IgG levels were depressed significantly in patients with stages A and B carcinoma of the prostate as compared to controls. Levels in patients with stages C and D lesions exceeded control levels but the difference was not statistically significant. Serum IgA levels in patients with stages A and B tumors were comparable to control levels but levels in patients with stages C and D lesions were significantly higher than controls.


Deep venous thrombosis following transurethral resection of the prostate: diagnosis by phleborheography. Deep venous thrombosis is a potential complication of transurethral resection of the prostate. We evaluated 150 paients undergoing transurethral resection of the prostate for benign and malignant disease to determine the postoperative incidence of deep vein thrombosis, using phleborheography as the instrument of detection. Phleborheography is an accurate, inexpensive, non-invasive method that uses low pressure transducers to detect volumetric changes in the lower extremity through recording cuffs. A 4.6 per cent incidence of deep venous thrombosis was detected by this technique. At the time this complication was discovered no patient exhibited clinical signs of thrombophlebitis, which reinforces the belief that clinical diagnosis alone is not a reliable screening technique for deep venous thrombosis. Anticoagulant therapy appears to be effective and safe in the treatment of this postoperative complication.


Prevention of bacteriuria after transurethral prostatectomy with nitrofurantoin macrocrystals. Of 40 patients who underwent transurethral prostatectomy without receiving antimicrobial chemoprophylaxis 13 had significant bacteriuria at some time in the postoperative course. Of 47 patients given nitrofurantoin none of the 42 for whom culture results were available had bacteriuria. This difference between the 2 groups is statistically significant (p less than 0.01). Nitrofurantoin prevents post-prostatectomy bacteriuria and has a theoretical advantage over agents such as the cephalosporins in that its action is limited to the urinary tract.


Herpesvirus type2: study of semen in male subjects with recurrent infections. Semen from 30 healthy male subjects with recurrent infections with herpesvirus type 2 was obtained when subjects were free of lesions and surveyed by tissue culture for an infectious virus in an attempt to elucidate the transmission of this disease. Inclusion bodies compatible with herpesvirus were found in tissue cultures of semen from 2 participants but an infectious virus could not be cultured directly from any sample. The data suggest that herpevirus type 2 is not ubiquitous in semen of male subjects with recurrent genital infections. The possible role of seminal inhibitors and a defective virus in causing the observed results is discussed, as are the current theories of herpesvirus type 2 transmission.


[Determination of pregnancy-associated glycoprotein for diagnosis and control of tumours in childhood]. Of 31 patients with solid, malignant tumours, 15 were shown to have over 0.5 mg/100 ml Serum concentration of alpha-2 pregnancy-associated glycoprotein (alpha2-PAG). These increased values were found for patients with neuroblastomas, Wilms tumours and rhabdomyosarcomas. However, for several different lymphomas and haemoblastoses, alpha2-PAG could not be demonstrated with the electroimmunodiffusion technique used (Laurell). From 230 children with non-neoplastic diseases, 62 had demonstrable concentrations of alpha2-PAG, of 38 healthy children, 2 were positive, while from 50 newborn infants, with different illnesses no increased values were found. Post-operative controls using repeated determination of d2-PAG are complicated by the slow change in concentration, and by the influence of chemotherapy. The determination of alpha2-PAG for the initial diagnosis of malignant diseases in childhood and for post-operative monitoring of the disease, is therefore of only limited value at least until further investigations have been carried out.


Limited cleavage of human immunoglobulins by elastase of human neutrophil polymorphonuclear granulocytes. Possible modulator of immune complex disease. Human polymorphonuclear lysosomal granules contain a neutral protease which is capable of digesting human immunoglobulins. The protease is located in the azurophil granules and enzymatically cleaves IgG, producing a Fab-like piece which is recoverable. The Fc-like fragment is not crystallized. The Fab-like fragment competes effectively for ligands formed by native antibody from which the fragments were produced.


Successful palliation using partial cardiopulmonary bypass in a two-day-old infant with type B interruption of the aortic arch. Partial cardiopulmonary bypass from the pulmonary artery to the descending aorta was used successfully for resuscitation and aortic arch reconstruction in a 2-day-old infant with type B interrupted aortic arch. Partial cardiopulmonary bypass interrupted the fatal progression to acidosis and death caused by hypoperfusion of structures distal to the ductus arteriosus in this "ductus dependent" infant, and this approach provided time for an accurate dissection and anastomosis. The natural history, pathophysiology, and currently available surgical alternatives are considered in suggesting a possible role for partial bypass in the treatment interrupted aortic arch.


Mast cells of the Mongolian gerbil (Meriones unguiculatus). Morphology, histamine content and role of calcium in the histamine release process. The fine structure, histamine conten;, and role of calcium in the histamine release process were studied in peritoneal mast cells of the Mongolian gerbil (Meriones unguiculatus). Stereological methods were applied to obtain quantitative data on their structure. The findings were compared with results obtained from the same type of cells in the rat. The gerbil mast cells were smaller in size (mean volume 242 micrometer3 vs 684 micrometer3 in the rat), and the nuclei were also smaller (55 micrometer3 vs 102 micrometer3). There were fewer granules in the gerbil mast cells and their diameter averaged 0.54 micrometer as compared with 0.78 micrometer in the rat). Only 20% of the cytoplasm of the gerbil mast cell was occupied by granules. This figure is approximately one third of that obtained in rat mast cells. The mean total histamine content per cell was 9 pg as compared to an estimated 30 pg/cell in rats. Calculated molar concentration of histamine in the mast granules, however, was higher in the gerbil than in the rat (2.3 M vs. 0.9 M). The mast cells of the gerbil were much more sensitive to the histamine-releasing agent compound 48/80 and in contrast to rat mast cells they were entirely dependent on calcium for their amine release. The fine cellular structure of both species showed multitudinous plasma membrane folds on their surfaces. In addition gerbil mast cells showed extensive surface invaginations. Apart from this were no major differences at the ultrastructural level between unstimulated cells of the two species. During histamine release, however, the mast cells of the gerbil showed a much greater tendency to form large, intracytoplasmic vacuoles and a decreased propensity for fusion of perigranular and plasma membranes (exocytosis) as compared with the corresponding cells in the rat.


Sensitisation to halothane-altered liver components in severe hepatic necrosis after halothane anaesthesia. In-vitro sensitisation (inhibiton or stimulation of leucocyte migration) in response to a liver homogenate obtained from rabbits pretreated with halothane was found in eight of twelve patients with halothane-associated hepatitis. Sensitisation was not observed when the homogenates were obtained from animals pretreated with ether. Furthermore, leucocyte migration in response to "halothane homogenate" was normal in eleven patients who had shown no abnormality in liver function after halothane anaesthesia and in thirty patients with other liver diseases. These studies provide direct evidence that sensitisation to halothane-altered liver-cell components is present in those occasional patients in whom severe liver damage develops after halothane anaesthesia.


N-acetyl-beta-D-glucosaminidase: A new approach to the screening of hypertensive patients for renal disease. Urinary activity of the enzyme, N-acetyl-beta-D-glucosaminidase (N.A.G.) was examined as a screening test for renal disease in patients with hypertension. Renal disease was present in 64% of patients with increased N.A.G., compared with only 14% of patients with normal N.A.G. The measurement of N.A.G. is cheap and convenient and this simple test warrants further assessment in hypertension as a guide to the need for further investigation.


Lipid-lowering effect of carnitine in patients with type-IV hyperlipoproteinaemia. Serum-lipid concentrations were determined in patients with type-IV hyperlipoproteinaemia treated with 900 mg/day oral DL-carnitine chloride. Serum-triglyceride was significantly reduced and concentrations continued to decline as carnitine administration continued. Total and esterified cholesterol concentrations did not change. Intravenous infusion of carnitine produced the same effects. The results suggest that carnitine is of value in the therapy of type-IV hyperliproteinaemia. Increased oxidation of free fatty acids in the tissues seems to account for the effects of carnitine on serum-lipid concentrations.


Alterations in human high-density lipoproteins, with or without increased plasma-cholesterol, induced by diets high in cholesterol. High-density lipoproteins (d=1.095--1.21) (H.D.L.( were isolated from six healthy men and women who added 4 to 6 eggs per day to their diet for 4 weeks and from five individuals who gradually increased their egg consumption to 3 per day over an 18-week period. Pre-diet and post-diet H.D.L.-binding activities for the cell-surface receptors of fibroblasts were compared by determining the quantity of 125I-labelled low-density lipoprotein which was competitively displaced by H.D.L. in binding, internalisation, and degradation assays. Irrespective of whether plasmacholesterol changed during the course of the diet, the binding activity of the post-diet H.D.L. was enhanced 2.6-fold to 4-fold compared with pre-diet activity. Furthermore, the increased binding activity in the H.D.L. could be accounted for by a minor, but potent, H.D.L. subfraction precipitated by heparin/manganese. Both binding activity and heparin precipitability appeared to correlate with an increase in arginine-rich apoprotein (apo-E) in the active H.D.L. subfraction. These data show that consumption of large numbers of eggs, whether or not it leads to an increase in plasma-cholesterol, does alter the properties of human H.D.L.


Microscopic examination of urine.

Microscopic examination of urine. Two methods of expressing microscopically observed leucocytes, erythrocytes, and casts in urine, both with centrifugation, one quantitatively (per ml) and the other per high-power field (H.P.F.), were compared for reliability in predicting renal functional abnormalities. In a series of 88 duplicate urines, abnormal numbers of casts or white or red blood-cells were detected by the quantitative method im 74% of patients with abnormal renal function but in only 36% by the N.P.F. method. The hourly excretion-rate of leucocytes over 3 1/2 h correlated very closely with the leucocyte concentration (cells/ml) measured by the quantitative method, so that a timed collection of urine was unnecessary. Unless findings from urinary microscopy can be relied upon, through adoption of a quantitative method, abnormal urines may be judged normal and some cases of glomerulonephritis may be falsely designated "clinically silent".


The MB isoenzyme of creatine kinase as an indicator of severity of myocardial ischaemia. 86 patients with ischaemic heart-disease were assessed for the presence of the MB isoenzyme of creatine kinase (C.K.-MB). Raised C.K.-MB levels with normal total C.K. were found in 10 patients with a clinical picture, but no electrocardiographic evidence, of acute myocardial infarction. 4 of these patients later had an infarction, but the remaining 6 have remained undiagnosed. Of 26 patients with unstable angina, 11 (42%) had high levels of C.K.-MB in the presence of normal total C.K. Of 50 patients with effort angina and a positive ergometric stress test, 3 (6%) with crescendo angina had high C.K.-MB levels in the presence of normal total C.K. In patients with angina, myocardial necrosis was excluded by normal serum-myoglobin levels. In both groups with angina, those with raised C.K.-MB levels had a more severe clinical picture, greater depressions of the ST segment, and lower threshold to ergometry. Raised C.K.-MB levels may indicate myocardial ischaemia.


Role of pineal gland in aetiology and treatment of breast cancer. The hypothesis that diminished function of the pineal gland may promote the development of breast cancer in human beings is suggested by the relation between breast cancer and prolonged oestrogen excess, and by the observation that the pineal secretion, melatonin, inhibits ovarian oestrogen production, pituitary gonadotrophin production, and sexual development and maturation. The hypothesis is supported by the following points. (1) Pineal calcification is commonest in countries with high rates of breast cancer and lowest in areas with a low incidence; the incidences of pineal calcification and of breast cancer are moderate among the black population in the United States. (2) Chlorpromazine raises serum-melatonin; there are reports that psychiatric patients taking chlorpromazine have a lower incidence of breast cancer. (3) Although information is lacking on breast cancer, the pineal and melatonin may influence tumour induction and growth in experimental animals. (4) The demonstration of a melatonin receptor in human ovary suggests a direct influence of this hormone on the ovarian function, and possibly oestrogen production. (5) Impaired pineal secretion is believed to be an important factor triggering puberty (early menarche is a risk factor for breast cancer).


Team approach to long-term intravenous feeding in patients with gastrointestinal disorders. A system has been developed for the prolonged administration of intravenous feeding in general wards. The service is coordinated by a clinician, specialist nurse, and pharmacist. Under local anaesthesia a silicone-rubber catheter is introduced through an infraclavicular incision to create a skin tunnel. By this technique 22 of 27 treatment periods (mean duration 42 days) required the use of only one catheter each. Ward care is simplified by the use of 3 litre bags containing aminoacids, glucose, electrolytes, and trace elements. The bags are prepared under laminar-flow conditions by a pharmacist in a specially designed room. Vitamins are added to the mixture and 'Intralipid" is infused separately as needed. In 38 treatment periods over a total of 1551 days weight-gain and gain in fat-free mass when required were satisfactory; the weight-gain correlated with increase in arm-muscle cross-sectional area (r = 0.72). Patients who have learnt to use the catheter have left the hospital for periods of up to 3 days, and 2 patients have given themselves intravenous supplements regularly at night in their homes.


Hepatitis "C" antigen in non-A, non-B post-transfusion hepatitis. Evidence for a new hepatitis-specific antigen has been obtained from double immunodiffusion assays between acute and convalescent sera obtained from patients with non-A, non-B post-transfusion hepatitis. The designation hepatitis C (HC) antigen is proposed. HC was found in the acute-phase sera of all 13 non-A, non-B post-transfusion hepatitis patients with longer incubation and duration periods (type 2) tested, but only transiently in 4 out of 10 acutephase sera obtained from patients with type 1 non-A, non-B hepatitis, with shorter incubation and duration periods. The antigen was also detected in 2 out of 16 single specimens obtained during the acute phase from acute hepatitis patients who had not received a blood-transfusion. This suggests presence of a carrier state. No patients with alcoholic hepatitis and no healthy blood-donor carried HC antigen. The antigen seems distinct from those of hepatitis A and B (surface and core). It migrated in the serum beta-globulin region and had a buoyant density of 1.30 and a molecular weight between 100 000 and 300 000. Antibodies against HC antigen were found in only 30% of the type-2 non-A, non-B post-transfusion hepatitis patients and did not persist for long. However, these antibodies were directed specifically against HC antigen and moved in a manner similar to 7S globulin on rate-zonal centrifugation.


Cholera, non-vibrio cholera, and stomach acid. Fasting and postprandial stomach acid production were low in 16 of 37 Bangalees convalescing from cholera or non-vibrio cholera. Gastric juice of hypochlorhydric patients did not kill cholera vibrios in vitro, whereas that from normochlorhydric patients rapidly killed vibrios in concentrations up to 10(10)/ml. To determine whether hypoacidity resulted from cholera or was a common predisposing factor, basal and betazole-hydrochloride-stimulated acid production were measured before and after cholera in a second group of patients consisting of American volunteers participating in a vaccine development programme. Cholera did not alter the stomach acid secretion of American volunteers, but low precholera basal acid production predispose to severe cholera. The results indicate that hypochlorhydria observed in convalescent Bangalee cholera patients is not caused by cholera, and must therefore have preceded it. Idiopathic tropical hypochlorhydria may be a major factor accounting for the high incidence of diarrhoea due to acid-sensitive pathogens in developing countries.


Cannabis, hypochlorhydria, and cholera.

Cannabis, hypochlorhydria, and cholera. In 90 volunteers participating in a vaccine-development programme consumption of beer more than 3 days a week was linked with high stomach acid output, and smoking of cannabis greater than 2 days a week was linked with low acid output. In 92 volunteers challenged with Vibrio cholerae or enterotoxigenic Escherichia coli, heavy use of cannabis was associated with more voluminous diarrhoea. Cannabis use may be an important factor predisposing to severe diarrhoea.


Controlled study of hypertransfusion during remission induction in childhood acute lymphocytic leukaemia. In a prospective, controlled trial 26 anaemic, neutropenic children with newly diagnosed acute lymphocytic leukaemia were randomised in pairs to receive either transfusion to a haemoglobin of 10--12 g/dl where clinically indicated (group A) or hypertransfusion to a haemoglobin of 16--18 g/dl (group B). Compared with group A (11 of 13 transfused), group B (all transfused) had a significantly more rapid rise in neutrophils at 7 and 10 days post-transfusion, a lower incidence of infection, and less interruption to chemotherapy. Hypertransfusion restored the myeloid/erythroid ratio to normal in bone-marrow of 5 of 6 children and the proportion of early myeloid precursors was greater than in controls.


Inhibitor of in-vitro granulopoiesis in plasma of patients with renal failure. Plasmas from 31 patients with moderately severe to severe renal failure inhibited granulopoietic colony formation in human marrow in vitro. The inhibitory activity could not be removed by in-vitro dialysis but was present in an ultrafiltered fraction of molecular weight less than 25 000 daltons. It inhibited the production of colony-stimulating activity (C.S.A.) by leucocytes in the culture system but had little or no effect on preformed C.S.A. or on the granulopoietic colony-forming cell itself. The level of plasma inhibitory activity correlated with the degree of azotaemia but not with the neutrophil or monocyte counts. Despite the potency with which granulopoiesis was inhibited in vitro, none of the patients was severely neutropenic, and only 4 had mild neutropenia (neutrophil count 1.5--2.1 x 10(9)/1).


A defect in catecholamine neurons in patients with prolactin-secreting pituitary adenoma. Although basal plasma concentrations of dopamine, noradrenaline, and adrenaline in patients with prolactin-secreting pituitary adenoma are normal, responses to oral bromocriptine are not. 1 and 2 h after bromocriptine, plasma concentrations of these three catecholamines are decreased in normal men but not in patients with hyperprolactinaemia secondary to prolactin-secreting pituitary adenoma or in normoprolactinaemic patients in whom a pituitary adenoma has been removed. Patients with prolactin-secreting pituitary adenoma probably have a defect in the regulation of brain catecholamine metabolism.


Haemolytic-uraemic syndrome: deficiency of plasma factor(s) regulating prostacyclin activity? It is suggested that patients with the haemolytic-uraemic syndrome and related disorders (such as thrombotic thrombocytopenic purpura) lack a plasma factor which stimulates prostacyclin (P.G.I2) activity. Normal plasma would supply the missing factor and is a rational treatment for some life-threatening symptoms (thrombocytopenia, haemolytic anaemia, hypertension) of this syndrome.


Radiotherapy and hyperbaric oxygen. Report of a Medical Research Council Working Party. The M.R.C. Working Party has coordinated randomised clinical trials to assess hyperbaric oxygen as a sensitiser in radiotherapy. 1669 patients were registered in these studies between 1963 and 1976. Hyperbaric oxygen significantly improved both survival and local tumour control after radiotherapy for head-and-neck tumours and for advanced carcinoma of the cervix. In carcinoma of the bronchus there seemed to be some improvement in survival but this was not statistically singificant. In carcinoma of the bladder hyperbaric oxygen has shown no benefit. Centres already equipped with hyperbaric chambers should continue to use them for those types of tumour shown to benefit. Since hyperbaric oxygen treatment makes great demands on medical and other staff, extension of its use must await comparison with other methods for improving radiotherapy which are now being evaluated.


Educating deaf and partially hearing children individually in ordinary schools. Research in North London and elsewhere has shown that nearly all hearing impaired children, when adequately supported, can be better educated in their local ordinary schools than in deaf schools or units.


Ribonucleic acid in the immune response. In the studies of experimental salmonellosis, immunization of mice with a live vaccine SER of S. enteritidis was found to be effective against further infection with virulent S. enteritidis 116--54. Macrophages obtained from the peritoneal cavity, subcutaneous tissue or liver of immunized mice inhibited intracellular growth of bacteria and resisted cell degeneration caused by engulfment of virulent 116--54 bacteria. This immunity was called cellular immunity. We discovered by chance in 1961 a transfer agent of immunity (TA) from the culture fluid of immunized macrophages. This agent is RNA in nature and can be extracted from the spleen, peritoneal exudate cells or the lymph node of immunized animals and is called immune (i) RNA. We could demonstrate antibody activity in macrophages treated in vitro or in vivo with iRNA by the immune adherence hemagglutination technique. Cellulr immunity against tumor cells could be transferred in vitro or in vivo to lymphocytes through iRNA prepared from the spleen cells of syngeneic, allogeneic and xenogeneic animals immunized with the tumor cells. We prepared iRNA against antigens capable of inducing humoral antibody production in animals, i.e., RBCs, bacterial toxin, bacterial flagella and hapten-protein conjugates. Serum antibody was not demonstrated in recipient animals of iRNA's by single or repeated injections of these agents. However, in these animals an increase in the number of specific antibody-carrying cells was found as rosette-formers. It was found further that prior injection of iRNA could induce immunologic memory and produced a high titer of humoral antibody after a boosting stimulation with a small dose of the corresponding antigen. The required interval between the first iRNA and the second antigenic stimulation, and the minimal effective doses of iRNA and antigen are described. We studied the interaction of iRNA with either T- or B-cells and with both cells using adoptive transfer system, athymic nude mice and neonatally thymectomized (NT) mice. Immune rna's against T-dependent and T-independent antigens could not induce the proliferation of antibody-carrying cells in cyclophosphamide-treated (B-cell depleted) mice. But these agents could induce the proliferation of rosette-formers, implying that iRNA's can replace some role of T-cells even against T dependent antigens. B-cells can be directly activated by treatment with iRNA against both T-dependent and T-independnet antigens, and they differentiated into rosette-formers. Passive transfers of iRNA were successful in establishing immunity against infection with S. enteritidis, or immunity to Salmonella flagella, RBCs and hapten-protein conjugates. The ability of iRNA to confer a secondary response of antibody formation is serially and passively transmissible in recipient animals. These facts suggest the presence of some mechanism that is responsible for the amplification of antigenic stimulation in the immune response...


[Electron microscopic structure of the mycelium of Actinomyces chrysomallus, the producer of the antibiotic chrysomallin, during the stab fermentation process]. Ultrastructural changes in the mycelium of Actinomyces (Streptomyces) chrysomallus 2703 producing the antibiotic chrysomallin in the process of submerged fermentation are described. When the antibiotic was produced at a high rate, most hyphae remained viable for a long period of time and had the ultrastructure typical of the young cells of actinomycetes. At the same time, a number of degenerative changes due to aging were observed; these resulted in autolysis. A decrease in the antibiotic titre was accompanied with intensive sporulation in a considerable number of the submerged hyphae. Structural changes in the sporeforming hyphae distinctly differed from those in the vegetative ones: the cell walls became thicker and numerous septa were formed. The exogenous formation of submerged spores in the culture under study has been studied in detail for the first time.


[The therapeutic importance of cimetidine (Tagamet) (author's transl)]. An overview of the development, actions and clinical applications of cimetidine, a new histamine H2 receptor antagonist, is given with reference to the literature. The experience of various investigators with this drug in the treatment of peptic ulcer disease are compared with each other and further indications described. The present results suggest a high therapeutic efficacy with trifling side effects. The latter is discussed with reference to two of our own observations. As long as the side effects are not sufficiently known, it is recommended that indications should be restricted to serious recurrent ulcer disease, reflux disease, prophylaxis and treatment of acute life-threatening hemorrhages and for palliative therapy of the Zollinger-Ellison syndrome.


Immunodiagnostics of malignant disease. VI. Electrophoretic mobility test (EMT) in malignant melanoma. The electrophoretic mobility test (EMT) is an in vitro assay for demonstrating cellular immunity. In the presence of tumor antigens lymphocytes of tumor patients liberate lymphokines, which reduce the charge of indicator particles resulting in a measurable reduction of their eletrophoretic mobility. Lymphocytes of 174 patients were tested by EMT. The antigens used were a basic myelin protein termed encephalitogenic factor (EF) and a 3M KCl extract from melanoma tissue. In 91% of the cancer patients there was a positive lymphocyte response. In contrast to this the controls and non-malignant diseases showed a positive result in only 8.7% of the cases. Using the 3M KCl extract from melanoma tissue as tissue as antigen 1 of the benign controls, 3 patients with nonmalignant diseases and none of the 49 patients with malignant diseases reacted positively, whereas in the melanoma group 86% showed a positive lymphocyte response. The results show the possibility of demonstrating tumor specific immune reaction in the EMT.


Antibodies to N6-(delta2-isopentenyl) adenosine and its nucleotide: interaction with purified tRNAs and with bases, nucleosides and nucleotides of the isopentenyladenosine family. The interaction of antibodies directed toward N6-(delta2-isopentenyl)adenosine, i6Ado, or its nucleotide with related bases, nucleosides, nucleotides and purified tRNAs is described. The selectivity of the antibody preparation was tested in inhibition experiments utilizing a sensitive radioimmunoassay to quantitate the binding of [3H]i6Ado to the antibody. Purified tRNAs containing various modified nucleosides adjacent to the 3'-end of the anticodon were tested to provide information about the selectivity of the antibody preparation toward nucleotides in this position of the tRNA chain. Antibodies directed against the nucleotide hapten were used to purify tRNAs which contain i6Ado and to quantitate the amount of that nucleotide. The same order of selectivity was expressed whether the nucleotides were free or in a tRNA molecule. Interaction of the antibody with compounds from the i6Ado family demonstrated dominance of the hydrophobic isopentenyl group and the importance of positional differences of modifications.


Hepatic function testing.

Hepatic function testing. The clinical situation determines the choice of hepatic function tests. Alkaline phosphatase (ALP) and aspartate aminotransferase (GOT) tests serve to detect disease, and when used in combination with a gammaglutamyl transferase (GGTP) test, to exclude it. The combination of ALP, GOT, bilirubin, lactate dehydrogenase (LDH), albumin, globulin, and GGTP tests is useful for routine differential diagnosis. Prothrombin time indicates severity of disease. Interpretation is facilitated by attention to ALP or GOT predominance; the relationship of LDH, ALP, and bilirubin; and the ratio of GGTP to ALP. Abnormalities on routine tests frequently do no more than point out the need for more definitive procedures.


3'-terminal nucleotide sequence of encephalomyocarditis virus RNA determined by reverse transcriptase and chain-terminating inhibitors. We have adapted the chain-termination method for determining the nucleotide sequence of DNA of Sanger, Nicklen, and Coulson [(1977) Proc. Natl. Acad. Sci. USA 74, 5463--5467] for use with reverse transcriptase (RNA-directed DNA nucleotidyltransferase) on RNA templates. With this method and using a primer (the octanucleotide pdT7rC) directed at the 3'-terminal poly(A), we have determined a sequence of 166 residues in the genomic RNA of the picornavirus encephalomyocarditis virus.


Ultrastructural antibody localization of alpha2-macroglobulin in membrane-limited vesicles in cultured cells. We have been developing a procedure for localizing intracellular antigens in cultured cells, by using peroxidase-labeled antibodies, that allows good morphologic preservation. Although useful, our previous technique did not preserve the morphology of membranes, and the location of the peroxidase reaction product was difficult to establish. In this paper, we report major improvements on the basic technique that markedly enhance the quality of localization and of morphology. Saponin is used to permeabilize membranes without destroying their morphology. The amount of reaction product is enhanced with a peroxidase-antiperoxidase label. The clarity of morphologic detail and contrast of reaction product density are increased by using postsectioning staining with the osmium/thiocarbohydrazide/osmium and uranyl acetate/lead citrate procedures. We have applied this technique to the ultrastructural localization of alpha2-macroglobulin and demonstrated that it is localized in membrane-limited vesicles. We have also used this method to improve the preservation of structures for localization by fluorescence microscopy.