Preparation of monospecific antiserums against porcine immunoglobulins, using agarose-linked immunosorbents. Immunoglobulins IgG, IgA, and IgM were isolated from porcine serum and milk, and antiserums against the 3 immunoglobulin classes were prepared. Monospecificity of the antiserums for the gamma-, alpha-, and mu-chains was obtained by absorbing them in agarose-linked immunosorbent columns. These immunosorbents were prepared by linking IgG or IgA-IgM to CNBr-activated agarose. Contaminating anti-alpha2-macroglobulin antibodies in the anti-IgA and anti-IgM serums were removed with agarose-linked fetal globulins.


Connective tissue-degrading enzymes of human leukocytes. Human leukocytes, when exposed to aggregated human gamma-globulin (AHGG) or immune complexes (isolated from RA synovial fluid) fixed to a cartilagenous surface, release neutral proteases that degrade the extracellular matrix of cartilage. The chondromucoprotein destruction by these proteases is suppressed by a variety of synovial fluids but is not susceptible to inhibition by trypsin, chymotrypsin, elastase inhibitors, or a combination of these agents. The inhibitory effect of synovial fluid can be reversed in the presence of increasing enzyme concentrations. Intact viable human polymorphonuclear leukocytes in the presence of AHGG also release a collagenase precursor that can be activated by limited proteolysis with trypsin or RA synovial fluids. Enzyme release (neutral proteases) by phagocytosing cells is inhibited by the antiinflammatory agents phenylbutazone and colchicine; these agents do not affect release of the collagenase precursor. However, the latent collagenase release is susceptible to inhibition when leukocytes are preincubated (prior to exposure to AHGG) with inhibitors of protein synthesis. Under these conditions, neutral protease release is unaffected.


Macrophage-mediated cytotoxicity: its possible role in rheumatoid arthritis. The association of macrophages with destructive processes in transplantation and tumoral, bacterial, and protozoan immunity illustrates their versatility in different situations. Their participation in inflammatory responses, including graft rejection and rheumatoid arthritis, would guarantee focal areas of high concentrations of hydrolytic enzymes and other macrophage products. That erosion of synovial tissue occurs in the presence of enzymes known to be associated with macrophages suggests that the large numbers of infiltrating macrophages and destruction of synovial tissue may be related.


Increase of melanocytes around malignant melanoma. In split epidermal sheets with clinically normal appearance a quantitative study was carried out on dopa-positive cells in the vicinity of malignant melanomas. These data were then compared with the number of melanocytes found in the skin of the contralateral body side of the same patient. In the epidermis around superficial spreading melanoma and lentigo maligna melanoma, the number of dopa-positive cells was usually significantly higher than in the contralateral body side. On the other hand, no difference was generally found around nodular melanoma.


Surgical treatment of ventricular arrhythmias using epicardial, transmural, and endocardial mapping. In 3 patients with ischemic coronary disease or primary myocardiopathy who were unresponsive to conventional and experimental antiarrhythmia therapy, surgical treatment of intractable ventricular tachycardia was performed using epicardial, transmural, and endocardial mapping techniques. An experimental canine model of acute and chronic myocardial ischemia with recurrent ventricular tachycardia was developed to refine the mapping technique for clinical use. In patients and animals alike, atrial overdrive pause pacing, premature ventricular pacing, or both were used to bring outa repeatable pattern of tachycardia. Mapping techniques distinguished the irritable focus so that surgical excision of the site of earliest activation could be performed. In addition, the mapping techniques were used in the validation following excision. The limitations of epicardial mapping alone in locating all areas of premature focus are discussed, and the need for mapping in ventricular aneurysm is demonstrated.


Clinical pathology of synovial fluid.

Clinical pathology of synovial fluid. Examination of synovial fluid should be performed in a systematic manner so as to derive the maximum of information. Synovial fluids should be divided into (1) non-inflammatory, (2) inflammatory, (3) purulent and (4) hemorrhagic types. In addition to general description, analysis should include mucin clot test, fibrin clog formation, microscopic examination for cell count and differential cell count, microscopic examination for crystals of gout and pseudogout and microgiological examination. Chemical examination should include estimation of glucose and uric acid. Immunochemical examination may include determination of immunoglobulins, antinuclear factor and LE factor.


Portable electrocardiographic monitoring: performance in patients with short P-R intervals without delta waves. Twenty-four-hour continuous protable tape-recorded electrocardiograms were obtained in 24 patients with short P-R intervals without delta waves. Atrial premature beats were noted in 15 patients (62%), paroxysmal supraventricular tachycardia (PSVT) in 5 (21%), ventricular premature beats in 14 (58%), and noticeable ventricular arrhythmia in 5 (21%). All episodes of PSVT reflected either unifocal or multifocal atrial ectopic firing. Atrioventricular nodal reentrant PSVT was not observed. Electrocardiographic correlation of symptoms with arrhythmias was not striking. In 21 of the patients, the P-R interval remained short constant through the 24-hour recording period. Patients with a short P-R interval without delta waves have frequent arrhythmias involving multiple areas of the conduction system. The presence of an accessory atrioventricular connection (James tract) would not explain the arrhythmias recorded in these patients.


Serum level of pregnancy associated alpha2-globulin in patients with spontaneous abortions. Pregnancy associated alpha2-globulin serum levels have been measured in abortion-cases. In cases of incomplete abortion with placental destruction and missed abortion this alpha2-globulin level significantly decreased, but no changes could be found when the placenta remained intact. The prognostic value of this pregnancy protein is discussed.


Studies of the thyroid function in children with Down's syndrome. The thyroid function (T-3 triiodothyronine; T-4 tetraiodothyronine and the captation of the I131 through the thyroid, in 2 and 24 hours after the intake of isotope radium), was studied in patients with Down's syndrome and the results being compared to those obtained from the respective siblings, all clinically normal. The comparative results showed no significant differences, except in the 2 hours captation of the I131 through the thyroid (P smaller than 0.01). The findings suggest that the hypothalamus-hypophysary or the response of the thyroid gland would be slow, but compensated when analyzed after 24 hours.


Angioneurotic edema. A review of modern concepts. We review the subject of angioneurotic edema with special emphasis on the more clearly defined entity of hereditary angioneurotic edema. The clinical presentations of the various forms of angioneurotic edema are discussed. Attention is directed toward the attempts that have been made to define this group of diseases in terms of their underlying biochemical mechanisms. A simplified schema of the serum complement system is presented. We conclude that only after we understand such biochemical interactions will we able to effect  a more definitive form of therapy for the angioneurotic edemas.


Proerdin and recurrent macroscopic haematuria. Kidney biopsies from ten children with recurrent macroscopic haematuria, showed mesangial deposition of IgG, IgA and complement. Eight of these ten biopsies also showed disposition of properdin, a basic euglobulin intimately involved in the alternate pathway of complement activation. Serum haemolytic complement activity was normal in the eight patients tested. Incubation of the serum at 4 degrees C for 24 dours did not result in any change in complement activity. Theses data suggest that the mesangial inflammatory process in these patients may be medicated in part by the alternate pathway of complement activation and that the mechanism is activated locally.


An evaluation of a new ECG monitor which detects and classifies ventricular extrasystoles. A device is described which is highly efficient in detecting and classifying ventricular ectopic beats.  This instrument may have a significant role in patient monitoring.  The system is based on analysis of a 5 electrode orthogonal system with comparison of QRS wave forms with a stored normal wave form.  The design was based on a test derived from 26 patients and evaluated further on long recordings of 16 other patients.  Results indicate that the device is highly reliable in QRS detection and in detection of ventricular ectopic beats.  The problem of artefacts due mainly to patient movement is held within resonable limits.


Characterization of murine sarcoma virus transformation of guinea pig cells and activation of an RNA tumor-like virus from nonproducer guinea pig cells. Guinea pig embryo (GEP) cells were transformed in vitro by the Kirsten strain of mouse sarcoma virus (Ki-MSV). The transformed cells were found to release infectious virus continuously and produced high titers of group-specific (gs) complement-fixing (CF) antigen characteristics of the murine sarcoma-leukemia virus complex. Foci of transformed cells were similar in appearance to those obtained with Ki-MSV in mouse and rat cells. The transformed cells produced RNA dependent DNA polymerase and type C virus particles with a density of approximately 1.15 g/ml in sucrose gradients by 3H-uridine labeling. The transformed cells produced tumors when transplanted into newborn guinea pigs. A number of focus-derived clonal lines from Ki-MSV transformed cells were isolated and characterized. All the focus-derived lines were found to be either producers or nonproducers (NP). The NP guinea pig cells produced neither infectious virus nor viral antigens of the murine sarcoma-leukemia virus complex although they were morphologically indistinguishable from virus-releasing MSV transformed GPE lines and produced tumors when transplanted into newborn guinea pigs. However, the sarcoma virus genome could be rescued in these NP cells by cocultivation with "helper" murine leukemia virus (MuLV) releasing GPE cells. Particles resembling guinea pig leukemia virus were activated from guinea pig NP cells or cultured normal guinea pig cells following chemical treatment. These particles were approximately 100 nm in the mature form and had a density of 1.16-1.17 g/ml. They contained RNA dependent DNA polymerase activity.


Progress in the chemotherapy of hematologic diseases. Progress in the chemotherapy of Hodgkin's disease and experimental therapeutic, pharmacologic, and clinical studies of the antitumor antibiotic, adriamycin, are presented in this abstract. In patients with disseminated Hodgkin's disease, the combination chemotherapy program (MOPP) produces a significant increase in the complete remission rate. This has been increased to 90% by the addition of low dose bleomycin to the MOPP program. The continuation of MOPP treatment beyond 6 months and to a total of 24 months provides improved results in patients in remission as measured either from time of onset of complete remission or from end of treatment. Finally, the pattern of relapse in patients with Hodgkin's disease provides a rationale basis for the selective use of radiotherapy in patients in complete remission. Adriamycin has a broad spectrum of antitumor activity in man. Its mechanism of action involves intercalation with DNA and inhibition of DNA function. The selective effect against tumors is not understood but may relate to membrane transport. Adriamycin is a highly important new antitumor agent for the treatment, not only of hematologic malignancies, but for a variety of solid tumors as well.


Possible cellular events in leukemia.

Possible cellular events in leukemia. An immunologically unresponsive state induced in experimental animals can be modulated by a number of procedures. Such modulation can result in the termination of the unresponsive state if there exists a normal complement of immunocompetent B cells in the presence of unresponsive T cells. It is possible to bypass the T cells and activate the B cells to respond to otherwise tolerated antigens. The bypass results from supplying a second (nonspecific) signal to the B cells, which, in the presence of antigen (specific signal), permits the B cell to differentiate. The role of the T and B cells and their activation and/or inactivation in the fate of both leukemic cells and oncogenic viruses is discussed.


Lymphomas and immunological tolerance in the rat induced by murine leukemia viruses. Several lines of Friend, Gross, and Rauscher virus-induced rat tumors were able to grow only in Friend, Gross, and Rauscher virus-tolerant rats that had received injections of each of these viruses at birth. When the tumors were transplanted into the virus-tolerant rats that had received injections of the other types of murine leukemia virus (MuLV), they grew initially and then regressed. Tumors did not grow in normal rats. The same is true in the methylcholanthrene-induced tumors that had been infected with MuLVs. It was deduced from these results that the transplantation antigen of tumor cells that were induced by infection with Friend, Gross, and Rauscher viruses may contain an antigen common to all three viruses and also an antigen that is individually specific, and that the host's immunological tolerance (induced by the neonatal injection of either Friend, Gross, or Rauscher virus) may consist of the tolerance common to all three viruses and, also, of the tolerance that is individually specific.


Sequences related to the RNA tumor viruses in the RNA and DNA of human leukemias and lymphomas. DNA-RNA hybridization was used to explore whether human neoplasias contain RNA molecules having sequence homologies to those of the RNA tumor viruses known to cause similar diseases in animals. The pattern of specific RNAs found in the human tumors showed a remarkable concordance with the predictions deducible from the animal systems. Thus human breast cancer contains RNA homologous only to that of the murine mammary tumor virus (MMTV). Human leukemias, sarcomas, and lymphomas (including Hodgkin's and Burkitt's) all contain RNA with sequence homology to the murine leukemia virus (RLV) and not to MMTV RNA. Finally, as in the case of the mouse, none of the human tumors examined contain RNA related in sequence to that of the avian myeloblastosis virus (AMV). The RNA detected in all of the human neoplasias was demonstrated to be of high molecular weight (1 times 10(7) daltons) and encapsulated with a reverse transcriptase in particles having densities between 1.16-1.19 g/ml. Further, the RNA of these human tumor particles was related in sequence to the murine viruses that cause the corresponding neoplasias in mice. Thus, 4 features diagnostic for the murine oncogenic viruses are satisfied by the particles found in the human cancers. Finally, it was shown by "recycling" experiments that the DNA from human leukemic cells and from lymphomatous tissue contained particle-related sequences that could not be detected in normal DNA. This finding was further substantiated by studies with identical twins in which it was shown that the leukemic twin contained particle-related sequences that could not be detected in the leukocytes of his identical healthy sibling. These findings are inconsistent with hypotheses that require chromosomal transmission in the germ line of complete copies of the information required to produce malignancy and the associated virus particles.


Transplantability, immunological unresponsiveness, and loss of cellular antigenicity in gross virus lymphoma. The capacity for syngeneic transplantation of rat lymphoma cells, originally induced by Gross lymphoma virus (GLV), was correlated with the expression of GLV-associated antigens by these cells. It was found that lymphoma cells that replicate GLV and display membrane and cytoplasmic GLV-associated antigens are consistently rejected when transplanted into syngeneic recipients. By contrast, lymphoma cells derived from the same parental cell line and morphologically indistinguishable, that have lost both surface and cytoplasmic antigenic expression were accepted and grew progressively at primary and metastatic sites. In further experiments, a long-lasting state of specific immunologic unresponsiveness was induced by administration of soluble GLV-associated antigens to newborn rats. These rats were later grafted with GLV-induced lymphoma cells that were positive for both membrane and cytoplasmic antigens. The rats conditioned at birth accepted the grafts which grew progressively, in contrast to normal controls which rejected them consistently. However, the grafted lymphoma cells showed progressive loss of antigenic determinants, and became serially transplantable in normal adult non conditioned rats. The membrane antigens were first to disappear from the grafted lymphoma cells, followed by the internal cytoplasmic antigens in further transplant generations. The transplantability of these cells increased accordingly. Titers of anti-GLV sera in tumor-bearing rats remained high even after surface antigens of lymphoma cells were no longer detected, but decreased to baseline levels after the internal antigens had been similarly lost.


Studies on the prevalence of type C virus associated with gibbon hematopoietic neoplasms. Gibbon malignancy frequently involves the hematopoietic system and can occur in clusters. Virus isolated from gibbon neoplasms possessed typical type C virus morphology, and the virion measured 100 nm in diameter with an electron-dense nucleoid measuring approximately 75 nm. The virus incorporated 3H-uridine into the nucleic acid and rested at a buoyant density of 1.14-1.16 g/cm3. Intra-and interspecific antigenic determinants were present, and the intraspecific antigenic determinant was shared with the woolly monkey sarcoma virus but not with feline or murine type C viruses. The virus and antibody reactive to the virus are more prevalent in gibbon groups that experience leukemia than those free of hematopoietic neoplasms.


Virus retrieval studies in human neoplasia. Short- and long-term co-cultures of 49 cases of human osteosarcoma cells with bone marrow or peripheral blood cells of patients with different types of leukemia were studied. Morphological changes were observed in 7 of 13 long-term co-cultures resembling those induced by RNA tumor viruses. The changes were accompanied by appearance of cytoplasmic antigen as shown by fixed immunofluorescence test with sera from patients with osteosarcoma, leukemia, and of some apparently normal blood donors. Absorption with Forssman-like substances, whole human embryo cells or osteosarcoma cells demonstrated the reaction to be due to tumor antigen(s) in co-culture cells showing morphological changes. Electron microscopy showed a few type C virus particles in one co-culture. Cell-free filtrates of fluid from the transformed co-cultures induced morphological changes in 1 of 4 human embryo cultures. Uninoculated embryo cultures or those inoculated with filtrates from parental sarcoma or leukemia cultures showed no morphological changes. Human embryo cell cultures treated with fluid from parental leukemic bone marrow but not from parental sarcoma cultures showed appearance of cytoplasmic antigen by immunofluorescence test with sera of osteosarcoma and leukemia patients and of some apparently normal blood donors. Transformed human co-cultures showed the cytoplasmic antigen with 28 of 48 sera of osteosarcoma and leukemia patients tested, after absorption with Forssman-like material, human embryo, and mycoplasma suspensions. Fourteen of 49 sera of normal donors were also positive with the transformed co-cultures. Similar results were obtained in an earlier series of experiments with human embryonic cultures transformed by fluid from different osteosarcoma-leukemia co-cultures when examined by fixed immunofluorescence tests with sera of patients with osteosarcoma and leukemia. In 2 whole human embryo cell cultures showing morphological changes high molecular weight RNA was found, similar to that of RNA animal tumor viruses and in one of the cultures transient reverse transcriptase was detected.


Detection and characterization of RNA tumor virus-specific DNA in cells. RNA tumor virus-specific DNA in cells can be detected by its capacity to 1) alter the reassociation kinetics of labeled double-stranded product of viral RNA-directed DNA polymerase; 2) anneal single-stranded DNA (cDNA) synthesized by viral polymerase; or 3) hybridize labeled viral 70S (genomic) RNA. Duplexes formed with these procedures can be analyzed for fidelity of base pairing, and the integration of viral DNA into the host genome can be established with a simple but stringent technique. We illustrate this methodology as applied to detection of Rous sarcoma virus (RSV)-specific DNA in XC cells and of mouse mammary tumor virus (MMTV)-specific DNA in murine and human tissues.


An endogenous oncornavirus of guinea pigs: its expression in leukemic cells. Guinea pigs do not have any known infectious oncornaviruses. We have recently activated an endogenous oncornavirus (GPV) from cultured guinea pig cells after bromodeoxyuridine riboside (BUdR) treatment. GPV is noninfectious for guinea pig cells and is inducible from any guinea pig cell in culture. Although the morphogenesis of the activated virus is unique, it has the morphology and density (1.16 g/ml) of type C oncornaviruses. It contains an oncornavirus specific reverse transcriptase and a high molecular weight RNA (70S) which dissociates into 36S following denaturation. GPV contains 5 major protein components (mol. wt. 95,000 to 16,000 daltons), 2 of which are glycoproteins. The viral genome which remains repressed in normal cells is expressed in leukemic guinea pig cells. These leukemic cells contain particles which have the morphological and biochemical characteristics of BUdR activated endogenous viruses. Unlike the cells transformed by infectious oncornaviruses, the leukemic cells contain the same amount of virus specific DNA present in normal cells.


Biochemical approaches to detection of viral related information in human acute leukemic cells. Two biochemical markers that have been utilized for the detection of viral related information in human acute leukemic cells are: 1) The reverse transcriptase and 2) a high molecular weight RNA with viral related nucleotide sequences. This paper summarizes evidence that shows that the reverse transcriptase isolated from human acute leukemic cells is biochemically related to the reverse transcriptase from RNA tumor viruses.


Evidence for an RNA tumor viruses in human milk. Certain human milks have been shown to contain particles that have the biochemical and biophysical properties that are diagnostic of the known RNA tumor viruses of animals. These properties include 1) a particle density of 1.16-1.19 g/ml 2) a viral reverse transcriptase (RNA-directed DNA polymerase), and 3) a high molecular weight (HMW) 60-70S RNA that contains polyadenylic regions of 200 nucleotides in length. Inner cores, or nucleoids, of these particles have been isolated. They have a density of 1.26-1.27 g/ml and contain the viral reverse transcriptase and 60-70S RNA. Using molecular hybridization, a specific homology was demonstrated between radioactive DNA synthesized from the RNA of the human milk particle and the RNA from human malignant breast tumors. RNA from benign breast tumors and other human tissues were negative in these tests.


Morphological, chemical, and antigenic organization of mammalian C-type viruses. New features in the architecture of mammalian type C viruses, in particular knoblike surface projections and hexagonally arranged subunits on the core shell could be demonstrated by electron microscopy, taking advantage of newly developed preparation techniques. As examples, murine leukemia viruses (MuLVs) and newly isolated porcine and bovine C viruses are presented. The major proteins of a MuLV were isolated and partially characterized in chemical terms and with respect to their serological and other biological activities, such as interfering and hemagglutinating (HA) capacity. Most of the characterized proteins could be localized in particular substructures of the virion either by selective removal or isolation of electron microscopically identifiable constituents. The information obtained allowed the design of a more detailed model of mammalian C viruses. Special attention was devoted to the further characterization of interspecies antigens of mammalian C viruses. Different antigenic determinants were revealed. Their distribution allows further subgrouping of mammalian C viruses.


Transcription of the Rous sarcoma virus genome in vitro and in vivo. RNA-directed DNA synthesis by detergent-disrupted virions of Rous sarcoma virus (RSV) initiates by the covalent attachment of pdA to the 3'-terminal rA of a 4S RNA hydrogen-bonded to the 70S RNA template. This 4S "primer" has structural features of tRNA and can be aminoacylated with methionine. Synthesis and integration of provirus DNA can be monitored in both permissive (duck) and nonpermissive (mouse) cells acutely infected with RSV. The results of these studies, as well as data obtained with RSV-infected mammalian cells which have reverted from a transformed to a pheno-typically normal state, indicate that integration of viral genes into the host chromosome is not sufficient cause for transformation. Pertinent features of virus-specific RNA-directed DNA synthesis in vitro and in vivo are reviewed and compared.


Evolutionary nature of human reverse transcriptase and of viral-related DNA synthesized in vitro by human leukemic cells. The reverse transcriptase and endogenous DNA product synthesized by virus-like particles in the cytoplasm of human leukemic cells have been studied for their genetic relatedness to homologous components obtained from several animal RNA tumor viruses. The human reverse transcriptase activity was inhibited by antibodies prepared against reverse transcriptase from some animal RNA tumor viruses. The DNA molecules synthesized endogenously by the human cytoplasmic particle in the presence of actinomycin D, using the reverse transcriptase enzyme and RNA template residing in the particle, hybridized to 70S RNA purified from certain animal RNA tumor viruses. Both the human reverse transcriptase and DNA product are closely related to homologues from primate type-C viruses, more distantly related to those from murine type-C viruses, and essentially unrelated to similar structures from feline or avian type-C viruses. They are not related to type-B RNA tumor viruses. The results demonstrate that the components from the human leukemic cells are viral (type-C) and primate in nature.


Further studies on the antigenic properties and distribution of the putative bovine leukemia virus. The C-type viruses found in long-term cultures. New Bolton Center (NBC) cell lines, of peripheral lymphocytes from leukemic cattle and in short-term cultures of bovine buffy coat(BC) cells share an immunofluorescent(IF)antigen detected in the cytoplasm of infected cells as well as an antigen demonstrable in gel diffusion experiments. Therefore the viruses from these cultures most likely represent different isolates of the putative bovine leukemia virus (BLV). The BLV precipitin antigen is analogous to the group specific (gs) antigens of the leukemia viruses of other species in that it is soluble, ether resistant, and apparently located within the virion. These observations, together with results showing that the specificity of the BLV precipitin antigen differs from that of the gs antigen of other mammalian leukemia viruses, indicate that the former antigen represents the intraspecies (gs-1) determinant of BLV. Antibodies to the precipitin viral antigen were found in 82% of cattle with leukemia and in 40% of clinically normal adult cattle in multiple-case herds. These groups of animals also had fluorescent antibodies to the virus, but with significantly higher frequencies (100% and 76%, respectively). On the other hand, in leukemia-free herds, precipitating antibodies were not found and the incidence of fluorescent antibodies was only 3%.


Comparison of drug effects on RNA tumor viruses and on transformed cells. Attempts to develop in vitro methods to test inhibitors for replication of C-type RNA tumor virus are made at two levels of the life cycle of the virus: 1) Formation of the proviral DNA, and 2) transcription and processing of viral RNA. Compounds such as rifamycin derivatives were found to be effective in interfering with the first process and compounds such as cordycepin, a nucleoside analogue, blocked effectively the second process. On the other hand, a class of known anti-tumor agents, glucocorticoids, was found to stimulate (10 fold or more) the induction of C-type virus following 5-iodo-2'-deoxyuride (IDU) treatment of murine cells. This is an intriguing example of a multi-focal action of an anti-tumor agent. We are exploring the implication of this for relapse or retransformation following successful chemotherapy in leukemic mice. Attempts are being made to develop an in vitro animal cell model for quantitative cytotoxic assays for compounds active in the above system and for comparing their effects on both normal and virus transformed cells.


[Effects of amantadine on heart and circulation]. The effect of amantadine-hydrochloride on heart and circulation is studied in 7 anesthetized, otherwise normal dogs with a mean body weight of 27.2 kg and in 8 heart-lung preparations of dogs. Arterial blood pressure, right atrial pressure, cardiac output and heart rate are measured and continuously monitored. Stroke volume and peripheral resistance are calculated. Left ventricular peak- and enddiastolic pressure, the rate of rise of intraventricular pressure and t-dp/dt are additionally measured in the heart-lung-preparations. Below 3 mg with kg(-1) in anesthetized dogs and 10 mg in the heart-lung preparation, respectively, a positive inotropic effect of amantadine is observed. This effect is caused by a liberation of catecholamines. Higher dosage of amantadine decreases cardiac contractility significantly. Therefore the negative inotropic influence of the drug itself has to be distinguished from the indirect sympathomimetic effect resulting from local release of myocardial catecholamines. Cardiac arrhythmias which occur in several experiments, can mostly be eliminated with propranolol or other drugs like lidocaine or sparteinsulphate.


Analysis of inhibition of lymphocyte cytotoxicity in human colon carcinoma. Serum inhibition of autochthonous lymphocyte cytotoxicity for tumour cells has been studied in 112 cases of colonic carcinoma. Addition of patient's serum to the lymphocyte tumour cell reaction mixture resulted in decreased cytotoxic reactivity of lymphocytes from 8 of 39 cytotoxic positive cases. It was also shown that sera could inhibit if separately preincubated with the lymphocytes (4 cases) or the target cells (2 cases). A tumour antigen preparation inhibited only when incubated with the lymphocytes. Inhibition by serum or antigen appeared to be specific for colon carcinoma. Four cases were specially studied to determine the mode of lymphocyte killing of tumour cells: in 3 it was mediated largely if not entirely by T lymphocytes, and in the fourth by both T and non-T cells. The findings support the view that T lymphocytes lose their anti-tumour reactivity in vivo in the presence of circulating antigen or antigen-antibody complexes such as would occur with progressive tumour growth.


Long-spacing collagen in skin biopsies from patients with lepromatous leprosy. In the course fo evaluating the effect of different drugs on the morphology of Mycobacterium leprae (Edwards, Draper & Draper, 1972) and on the phagocytic cells containing these organisms (Edwards, 1973), an unusual feature was observed in the dermal connective tissue of skin biopsies obtained from some of the patients. Variable amounts of an extracellular, cross-striated osmiophilic material were present in the dermis. The purpose of this communication is to describe the banded substance observed in the extracellular space of the lepromatous skin and to compare it briefly with similar substances and structures reported to occur elsewhere.


Preparation and characterisation of ribonuclease from human hypertrophic prostate gland (RNAase P2). An endo-type, cyclising, 3'-phosphate-forming rebonuclease was purified to homogeneity from a water/Tween 80 extract of human hypertrophic prostate gland. The enzyme is acid- and heat- resistant and is optimally active at pH 7.0, 0.1 M NaCl. Molecular weight determined by gel filtration on Sephadex G-75 and sucrose density gradient centrifugation gave a mean value of 15 000. The prostatic ribonuclease is inhibited by Cu2+, bromoacetate and photooxidation in the presence of methylene blue. Other divalent ions, EDTA and p-chloromercuribenzoate have no influence on the enzymic activity. Prostatic RNase resembles RNase A in that it preferentially cleaves linkages in RNA after pyrimidine nucleotides to produce oligonucleotides terminated in cyclic 2',3' phosphate. The enzyme is inactive with poly(A) - poly(U) as substrate. Poly(U) is hydrolyzed four times as fast as poly(C), and 1.2 times as fast as RNA.


Affinity chromatography of galactose containing biopolymers using covalently coupled Ricinus communis lectin to Sepharose 4B. A galactose-specific lectin isolated from Ricinus communis beans has been covalently coupled to Sepharose 4B activated with cyanogen bromide. The immobilized lectin retains its polysaccharide-binding property. The Sepharose-lectin can be used for the purification of polysaccharides containing terminal nonreducing galactose. Only a small fraction of 'native fetuin' and 'native ceruloplasmin' are retarded on Sepharose-lectin. On analysis it was observed that they had a lower content of sialic acids as compared to the native and unbound glycoproteins (sialated fractions). However, on desialation, fetuin and ceruloplasmin were completely adsorbed to Sepharose-lectin. The asialoglycoproteins interact strongly with Sepharose-lectin as compared to 'partially sialated glycoproteins'. This has been attributed to the exposure of galactose residues of these glycoproteins on enzymatic desialation. These experiments demonstrated that Sepharose-lectin interacts with glycoproteins through their terminal, non-reducing galactose. On the basis of these experiments it is suggested that Sepharose-lectin can be used as an analytical tool for separation of 'fully sialated glycoproteins' from the 'partially sialated glycoproteins'.


Solution behavior, circular dichroism and 22 HMz PMR studies of the bovine myelin basic protein. Bovine myelin basic protein has been investigated with regard to its solution behavior, circular dichroism and 220 MHz PMR spectral properties. At pH 4.8 gamma/2=0.1 acetate buffer, light scattering yielded a Mr of 17 700 and a virial coefficient of 1.0-10(-4) mol-ml/g2. Above pH 7.0 the protein was found to aggregate to higher mol. wt species. Sedimentation experiments at pH 4.8 yielded s degrees 20,w of 1.27 S at gamma/2=0.1 and 1.46 S at gamma/2=0.35. The diffusion coefficient determined from ultracentrifugal experiments was 7.25-10(-7) cm2/s at gamma/2=0.1 and 0.35. The value of f/f0 from diffusion at pH 4.8 and gamma/2=0.35 was 1.64, corresponding to an axial ratio of 11 to 1. The radius of gyration was calculated as 4.28 nm and the root mean square end to end distance was 10.5 nm. At pH 9.0, gamma/2=0.1, s degrees 20,w was 1.71 S and D degrees 20,w was estimated at 7.4-10(-7) cm2/s. The behavior at pH 9.0 reverted to the behavior at pH 4.8 when the pH was readjusted. The E1%/1cm=5.64 at 276.4 nm and 225 at 196 nm. Titration of the protein with trifluoroethanol elicited three distinct regions of conformation stability having increasing helical content as the mol fraction of trifluoroethanol increased. The results of the present study have permitted some comparison of analogous properties and conformational behavior with the basic membrane protein cytochrome c.


Consecutive cytochemical staining for the analysis of the blastic population in the acute phase of chronic myeloid leukemia. Blastic crisis in chronic myeloid leukemia is characterized by several cytological alterations which may represent some abortive attempts to differentiate along various cell lines as a consequence of a maturation defect of the myelopoietic cells. These changes of the hematological picture are associated with alterations of the karyotype and with cytochemical abnormalities of the blast cells, possibly related to their metabolic anomalies. In this regard 14 patients with blastic crisis were investigated to achieve an evaluation of the composition of the cell population during the acute phase. A sequence of three cytochemical reactions applied consecutively on the same slide (alpha-naphthyl-acetate esterase + AS D-chloro-acetate esterase + PAS) proved to be useful for the detection of differently oriented blast cells. During the acute phase of chronic myeloid leukemia only about one half of the blast cells were expressing granuloblastic differentiation. The data may be important for some clinical and prognostic factors, since the heterogeneity of the blastic population may be associated with a particular resistance to therapy.


Fluorescent labeling of human platelets. Noncovalently bound fluorescent probes have been used to study changes in the platelet which may occur during platelet aggregation. Platelets were exposed to either N-phenyl-naphthylamine (NPN) or 8-anilino-1-naphthalene-sulfonic acid (ANS). Both dyes were bound by the platelet, and platelet aggregation by collagen or thrombin was unaffected by the presence of the label. No change in fluorescence intensity or wavelength of maximum intensity was observed during platelet aggregation.


Deoxycytidine kinase: properties of the enzyme from human leukemic granulocytes. Deoxycytidine kinase, which phosphorylates deoxycytidine (CdR) and its analog, cytosine arabinoside (ara-C), has been purified 71-fold from human leukemic cells. Biochemical properties of the partially purified enzyme included a molecular weight of 68,000, Kms of 7.8 muM for CdR and 25.6 muM for ara-C, and optimal activity with ATP and GTP as phosphate donors. Ara-C phosphorylation was strongly inhibited by CdR (Ki = 0.17 muM) and dCTP (Ki = 7.3 muM) and was weakly inhibited by ara-CTP (Ki = 0.13 mM). Purification by calcium phosphate gel elution and DEAE chromatography effectively separated this enzyme from cytidine deaminase, which deaminates both CdR and ara-C, and from uridine-cytidine kinase, the enzyme which phosphorylates 5-azacytidine. CdR kinase activity was found to decrease and cytidine deaminase to increase with maturation of normal and leukemic granulocytes. Myeloblasts purified by Ficoll sedimentation revealed an average kinase activity of 15.4 U/mg protein in acute myelocytic leukemia and 12.3 U/mg protein in blastic crisis of chronic myelocytic leukemia (CML). The average ratio of CdR kinase to deaminase activity in crude cell extracts varied from 0.197 in AML and 0.089 in blastic crisis to 0.0004 in normal granulocytes, reflecting the changes which take place with cellular maturation. The absolute levels of kinase and deaminase and the ratio of these two enzymes varied considerably among patients with AML, indicating that quantitative differences may be found in the metabolism of CdR and its analogs in leukemic cells.


Fluid hexagonal lattices of sub-units in negative staining preparations of fragmented human lymphocytes. Electron microscopic studies of negative staining preparations of fragmented human lymphocytes have revealed highly-ordered fluid hexagonal lattices of repeating 100 A sub-units with central depressions. It is supposed that the particles came from the plasma membrane of the lymphocytes.


Whither thyrotoxicosis?

Whither thyrotoxicosis? For 30 years the thyrotoxic patient has been subjected to a plurality of treatments by surgery, radio-iodine and long term anti-thyroid drugs. These therapies have been accepted as complementary to the needs of the individual patient, without regard for long term results or the economic situation as it affects both patient and hospital services. In the context of surgical treatment which is now available, it is suggested that the advantages of operation over other therapies presage a reversion to surgery as the treatment of choice.


Surgical treatment of thyrotoxicosis: results of 272 operations with special reference to preoperative treatment with anti-thyroid drugs and L-thyroxine. From 1959 to 1970, 272 operations for thyrotoxicosis were performed. Most of the patients received anti-thyroid drugs and thyroid hormones preoperatively. The patients were continuously followed up. The primary results with low morbidity and no mortality as well as the long term results with a low rate of recurrence and a relatively high incidence of thyroid substitution are discussed. A safe and effective programme for surgical treatment of thyrotoxicosis is described. Anti-thyroid drugs and thyroid hormones should be administered as the method of choice in preparing these patients for surgery.


Parathyroid identification by methylene blue infusion. The intravenous infusion of methylene blue was investigated as a procedure that would identify parathyroid glands during operation in 17 patients with primary hyperparathyroidism. The dye was found to stain all adenomas, most hyperplastic glands and occasionally normal parathyrodi glands. In addition, the frequency of glandular staining was directly related to the size of the gland, although size and histology may have been independent variables. Methylene blue infusion is a safe effective method of localizing abnormal parathyroid glands.


Isolation and characterization of an aetiological agent in Whipple's disease. A cell wall deficient form of an alpha-haemolytic streptococcus was grown from a prolonged monolayer cell culture of a lymph node taken from a patient with Whipple's disease. Serological cross reactivity was shown between the organism and the material within Whipple's disease macrophages positive for diastase-resistant periodic acid-Schiff (D./P.A.S.). In vitro studies characterized the organism as a facultative intracellular parasite which caused the accumulation within cells of D./P.A.S.-positive material. These results suggest that a pathogenic bacterium is the essential aetiological agent and that the culture of Whipple's disease tissues in hypertonic media may have practical value.


Treatment of superficial thrombophlebitis: a randomized, bouble-blind trial of heparinoid cream. In a prospective, double-blind, randomized trial the efficacy of a heparinoid in ointment form was assessed in treating superficial thrombophlebitis developing after continuous intravenous infusion. One hundred surgical patients were studied, and clinical examination and the iodine-125-labelled fibrinogen test used to assess the results. The mean time required for the relief of local symptoms and signs and the rate of local decline in radioactivity differed significantly between patients receiving the heparinoid cream and those recieving the placebo.


Low molecular weight specific proteins in identified molluscan neurons. II. Processing, turnover, and transport. Three identified neurons (R14, R15, and L2-6) from Aplysia californica synthesize specific, low molecular weight proteins which are further processed or converted into smaller sized specific proteins. These specific proteins have differential turnover rates, and appear to be selectively transported out of the individual neuronal somata at different rates. The latter transport process can be blocked by colchicine, a well-known blocker of axonal transport. The relationship of these phenomena to the functional activity of the individual neurons is discussed.


Anti-synaptic antibody in allergic encephalomyelitis. I. Neurophysiological studies, in guinea pigs, on the exposed cerebral cortex and peripheral nerves, following immunological challenges with myelin and synaptosomes. The potential evoked at the surface of the cerebral cortex of a guinea pig, by stimulation of the contralateral forepaw, usually consists of an initial double positive wave whose waveform remains unchanged during rapid stimulation. In a guinea pig with experimental allergic encephalomyelitis (EAE) the response is attenuated at low frequencies of stimulation. Observations were also made on animals with experimental allergic neuritis (AEN). These animals showed a peripheral specificity for decreased conduction velocity, but have normal cortical evoked responses. Histological studies were undertaken in parallel with the electrophysiological studies and the concordance and discordance between the two is discussed. We conclude that there is no evident correlation between the severity of the electrophysiological effects and the histological lesions. An attempt was also made to induce an immunological challenge in guinea pigs, in the same way that EAE and EAN is produced, by the injection of synaptosomes. No clinical signs or alterations in the histology or electrophysiology of the animals were seen. A discussion is included on the elucidation of the site of action of EAE by discriminating between the direct effects on myelin and synapses and by the indirect effects of myelin damage on synapses. No firm conclusion is reached and the matter is left for further analysis in the subsequent paper. Finally, the neurophysiological alterations, demonstrated in the EAE and EAN situations, are discussed in terms of a humoral factor possibly acting on the myelin sheath and indirectly affecting synaptic function. This matter is further discussed in the subsequent paper.


Anti-synaptic antibody in allergic encephalomyelitis. II. The synapse-blocking effects in tissue culture of demyelinating sera from experimental allergic encephalomyelitis. It has been suggested that demyelination cannot account for all of the observed clinical symptoms of multiple sclerosis (MS), in particular the rapidity of onset and remission of the disease, and attention has been focussed on the role of the synapse in 'demyelinating diseases'. In the present paper we have attempted to resolve the fundamental question of the site of action of a demyelinating disease, experimental allergic encephalomyelitis (EAE), by the use of cultures of neonatal rat cerebellum. Electrophysiological and morphological development in these cultures run hand-in-hand, and in the first few days in vitro there is a 4-5 day period when synapses are both seen ultrastructurally and known to be functioning but before the onset of myelination. The serum from guinea pigs with EAE was added to these cultures at different stages during their development and the morphological and electrophysiological effects observed. An abolitionary effect on the bioelectric activity of the culture was only observed when the serum was added to mature, myelinated cultures. Also the same active sera had no effect on synaptic activity before myelination had occurred. We conclude that the synaptic blocking effect occurs only when myelin is destroyed.


The localization of the basic protein and N-2 in diseased myelin. A non-penetrating reagent 4,4'-diisothiocyano-2,2'-ditritiostilbene disulfonic acid ([3H]DIDS) has been used to label isolated normal and diseased myelin. The basic protein and the hydrophobic protein, N-2, were isolated from each myelin. When normal myelin was labeled the specific activity of the basic protein was about 25% of that of the hydrophobic protein (N-2). The specific activities of these two proteins isolated from chronic multiple sclerosis myelin were similar to those of the normal myelin. In contrast, the specific acitivity of the basic protein isolated from acute multiple sclerosis myelin was about 400% higher than that of the basic protein isolated from either normal or chronic multiple sclerosis myelins. The specific activity of the N-2 protein was only 50% of that of the N-2 protein isolated from normal and chronic multiple sclerosis myelins. It was concluded that the arrangement of proteins in isolated chronic multiple scerosis myelin was not markedly altered in comparison to that of isolated normal myelin. However, the arrangement of proteins in acute multiple sclerosis myelin appeared to be considerably different from that of the other two myelins.


Axoplasmic transport in the toad Bufo marinus. The rate and course of axoplasmic transport from the eighth dorsal root ganglion cell bodies into the sciatic nerve of the toad Bufo marinus were studied. Concentrated tritiated proline was hydraulically injected into a surgically exposed dorsal root ganglion of animals maintained at 19 +/- 0.5 degrees C. At postinjection intervals of 1, 6, and 10 h, the animals were sacrificed and the dorsal root, ganglion, and sciatic nerve were removed bilaterally. The dorsal roots and peripheral nerves were cut into 3 mm segments measured from the ganglion. In some experiments all tissues were prepared for liquid scintillation counting techniques. In others the ganglion and every fifth 3 mm nerve segment were fixed in Bouin's fixative for radioautography, and the remaining tissue segments were prepared for liquid scintillation counting methods. Scintillation counts (counts/min) of consecutive segments along the labeled nerve were plotted against distance (mm) for each animal. Examination of these profiles showed a peak of radioactivity in the injected ganglion for each animal that was followed distally by an abrupt drop in the adjacent segments of the nerve. Radioactivity remained relatively stable in subsequent segments forming a plateau and then dropped to baseline levels forming a wavefront in the distal portion of the peripheral nerves of the 6 and 10 h toads. Movement of this wavefront during the 6 to 10 h time interval provided evidence for an axoplasmic flow rate of about 120 mm/day. Radioautographs of the ganglion and representative segments along the sciatic nerve were examined with both bright- and dark-field microscopy. Accumulations of silver grains were observed overlying the injected ganglion cell bodies and labeled axons of the nerve. Values for the number of silver grains/unit area were obtained from dark-field radioautographs of the nerve segments through the use of a computer-microscope system. The relative amount of radioactivity present in the axons was thereby demonstrated. Radioautographic data confirmed the axoplasmic flow rate of about 120 mm/day determined by liquid scintillation counting methods for the bulk of radioactive materials transported at this fast rate. In addition, the evaluation of radioautographic data suggests that an even faster flow rate of 185-215 mm/day may exist for a small portion of the labeled materials transported in the axon.


Slow axoplasmic transport of mitochondria (MAO) and lactic dehydrogenase in mammalian nerve fibers. The axoplasmic transport of the enzyme monoamine oxidase (MAO) (monoamine:O2 oxidoreductase, (deaminating) EC 1.4.3.4), a marker for mitochondria, and lactic dehydrogenase (LDH) (L-lactate:NAD oxidoreductase, EC 1.1.1927), a soluble component of axoplasm, was studied in cat sciatic nerve. For both these enzymes a linear accumulation was found in the nerve proximal to ligations over a period of at least 20 h. In double-ligation experiments no evidence of a depletion of enzymes within the nerve segment was found over this period of time as would be the case if some portion of the enzymes was carried by fast axoplasmic transport. Both the soluble protein enzyme LDH and the mitochondria, shown by MAO, are thus considered to be moved down the nerve by slow axoplasmic transport. Some differences in the two materials were seen in the greater fall in the level of MAO compared to LDH within the double-ligated segment over the succeeding period from 20 to 48 h. These changes are considered with respect to the transport filament model as modified to take into account slow axoplasmic transport.


Association of proteins undergoing slow axonal transport with goldfish visual system myelin. [3H]Proline derived radioactivity which is slowly transported in the goldfish optic nerve has been isolated in association with purified preparations of optic tectal myelin. Acrylamide gel analysis reveals that most of the slowly transported radioactivity is distributed among the high molecular weight myelin proteins with a major peak comigrating with a band (approximately 42,000 daltons) which is prominent in goldfish brain myelin. A similar peak is not seen in gels of whole synaptosome protein. Relatively small but measureable amounts of radioactivity comigrate with the basic and proteolipid proteins. A low density myelin subfraction is 3-5 fold lower in specific radioactivity (disint./min/mug protein) than a corresponding high density fraction and shows particularly low specific radioactivities in the basic and proteolipid bands. The results are discussed with reference to the possible adherence of specific axonal membrane fragments as well as other potential sources of myelin label.


The influence of complement on the neutralization of infectious bovine rhinotracheitis virus by globulins derived from early and late bovine antisera. Calves were inoculated at four week intervals with infectious bovine rhinotracheitis virus (IBRV). Sera were obtained at eight days (early sample) and 21 days (late sample) after each inoculation. Kinetic neutralization was carried out with 7S and 19S globulins derived from these sera, both in the presence and in the absence of guinea pig complement (C). In all instances the 19S neutralizing antibody (AB) was dependent on C for neutralization of IBVR. However, C dependency was not observed with any of the 7S preparations. Neutralizing activity was readily detected in 19S globulins from the early sera after primary and subsequent inoculations but not in any of the late sera. Early sera collected after primary inoculation did not contain any 7S neutralizing AB but it was present in all the other sera tested. The 7S AB when present was always at a considerably higher concentration than 19S AB. Thus it may be possible to determine whether cattle have recently been exposed to IBRV when paired serum samples are not available by determining the presence of C-dependent 19S globulins. In addition, by comparing 19S and 7S levels, a distinction may be made between primary and secondary responses to IBRV.


A scanning electron microscope study of human cerebral arteries. Human cerebral arteries were obtained from autopsy, fixed under pressure, cut open, and tacked onto pieces of cork. For one artery the intima was partly teased away, exposing the media, and treated with a silver nitrate process. For another artery the adventitia was exposed. Both arteries were processed through graded ethanols and coated with gold paladium for the scanning electron microscope. The collagen fibers of the adventitia were approximately 5 mum in diameter and consisted of a bundle of microfilaments, each of which had a diameter of 800-1000 A (1 A = 10(-10) m). The collagen fibers were oriented parallel to the long axis of the artery. The muscle cells of the media had a diameter of 2-5 mum and were arranged circumferentially with a pitch of approximately 20 degrees. The collagen fibers of the media travel perpendicular to the muscle cells, and parallel to the long axis of the artery. The fibrillar components of the elastin in the intima had a diameter of approximately 700-1000 A and were arranged parallel to the long axis of the artery. It was postulated that the fibrillar part of the elastin was the elastic component of the elastin.


Induction of avirulent variants in Erwinia stewartii by incubation at supraoptimal temperatures. High temperatures (37 degrees C) induced non-pigmented, and (or) small colony variants in some Erwinia stewartii strains. The former differed from the parent strain serologically and in having lost virulence to Zea mays. The small colony variants retained phytopathogenicity.


Analysis of soluble melanoma cell membrane antigens in metastatic cells of various organs and further studies of antigens present in primary melanoma. In malignant melanoma, using Sephadex G-200 chromatography and polyacrylamide gel electrophoresis (PAGE), it has been possible to separate two types of skin reactive antigens. The first, found in Sephadex fraction II and PAGE region a appears specific for melanoma. Allogeneic extracts have produced positive reactions in many patients with skin or ocular melanoma, and have given negative reactions in patients with other types of cancer or in patients with ocular lesions simulating melanoma. The second group of antigens, in Sephadex fraction III and PAGE region b were less specific. These antigens produced positive skin reactions in some patients with breast cancer, as well as in patients with melanoma. Reactivity to PAGE region a appeared to be confined to one protein band, but three different bands in region b gave positive reactions. A study was made of the presence or absence of similar antigens in metastatic deposits of malignant melanoma. Metastatic lesions in the following tissues were analyzed: liver, lung, adrenal, skin, and colon. These were compared with pooled primary skin melanomas by skin testing in the same patients. The tumor-associated melanoma antigen, found in Sephadex fraction II and PAGE region a appeared to be strongest in adrenal, lung, and liver metastases. It was found that the protein yield in this region was not indicative of the strength of the antigen. Therefore, a careful, detailed analysis of the protein bands present in PAGE regions a and b from primary skin melanoma was conducted. Only one band in PAGE region a was found to be responsible for positive skin reactivity. This band was found to be a glycolipoprotein. Further studies were also conducted in order to determine whether or not some of the antigens present might be fetal antigens. Some of the protein bands present in Sephadex fraction III and PAGE region b of melanoma appeared to be similar to some of the PAGE region b proteins present in fetal skin cells. Two bands from fetal skin also had the same location on PAGE as two bands from ductal breast cancer, although the relationship to melanoma region b antigens was not exact. These fetal proteins, which seemed to be present both in ductal breast cancer cell membranes and in melanoma cell membranes, might account for the positive skin reactivity seen in this region, and also for the cross reactivity of skin tests with this antigen.


Enzymatic modification of the transplant antigens (ABO system) on human tumor cells. Blood group glycosyltransferases were used to modify HeLa cells of H specificity (O Group) into cells of A and B specificity. We also obtained the identical type of modification with lymphocytes from healthy subjects and leukemia patients. This method can be applied to tumor cells in general, and constitutes an attempt to stimulate the immunocompetent system.


Bronchial carcinoid tumors.

Bronchial carcinoid tumors. Twenty-eight pulmonary carcinoid tumors were reviewed histologically and clinically. Hematoxylin-and-eosin-stained sections were utilized, as well as special stains, including the argyrophil and argentaffin reactions. The 22 tumors located centrally, at the level of primary or segmental bronchi, had a microscopic appearance distinct from those located more peripherally. One peripheral tumor that was large in size appeared much more aggressive histologically, and was designated an atypical carcinoid. The origin of carcinoid tumors from Kulchitsky cells in the lung, the distinction of peripheral tumors from chemodectomas, and the relationship of bronchial carcinoids to bronchial epithelial hyperplasias and oat cell carcinomas are discussed.


The function of fat bodies in relation to the hypothalamo-hypophyseal-gonadal axis in the frog, Rana esculenta. In this study the authors have tried to furnish experimental support for the importance of fat bodies in the normal functioning of the hypothalamo-hypophyseal-gonadal system of the male frog, Rana esculenta. These experiments have shown a hypothalamo-hypophyseal control of the mobilization of fat body contents, directly involved in the control of testicular activity. Furthermore it is proposed that the fat body contents are released into the testis through direct vascular contacts between the two organs. We suggest that the A1 cells (lactotrophs) and/or B2 cells (FSH-gonadotrops) of the pars distalis gonadotropins are incapable of stimulating the testis in the absence of fat bodies. In the light of these results a scheme has been put forward showing the position of fat bodies in the hypothalamo-hypophyseal-gonadal axis of the frog.


The distribution of oxidizable mucosubstances and polysaccharides in the planarian Polycelis tenuis Iijima. A chromic acid oxidation-silver technique was used to localize polysaccharide material in Polycelis tenuis at the electron microscope level. In the epithelium, staining was observed within apical vacuoles and on the free surfaces of the cells. A similar staining was observed in relation to the glycocalyx of the pharyngeal epithelia and that of the flame cells. Silver was deposited in the basement membrane. In the parenchyma, the major components giving a positive reaction were the cyanophil and mucous gland cells. Particularly strong silver staining (confirmed by X-ray microanalysis) was observed in the granules and Golgi apparatus of the cyanophil cells. IDPase activity was also found in relation to the Golgi apparatus and its secretory products. The overall distribution of mucopolysaccharide material was confirmed with the PAS and Alcian blue techniques. The fine structural localization of the Alcian blue was also determined using electron microscopy and X-ray microanalysis.


Dynamic properties of axonal transport of proteins and glycoproteins: a study based on the effects of metaphase blocking drugs in the developing optic pathway of chick embryos. Some properties of the axonal transport of proteins and glycoproteins along the optic pathway of chick embryos and newly hatched chicks were studied by labelling retinal ganglion cells with 3H-proline or 3H-fucose. A study of the effects of colchicine (COL) and vinblastine (VLB) on embryonic axonal transport was also carried out. Marked changes in the efficiency of axonal transport were found throughout development. In particular, the fraction of retinal ganglion cell proteins which is rapidly exported toward tectal terminals increases during embryonic life but steadily decreases after hatching. Glycoprotein transport behaves similarly except that its efficiency is relatively higher at stages when critical events of synaptic maturation in the tectum are reported to occur. Embryonic axonal transport is blocked by COL and VLB at very low intravitreal concentrations. Retinal protein synthesis and the morphology of ganglion cells are profoundly altered by the drugs: in general, COL and VLB effects were much more marked in embryonic than in mature neurons. An analysis of the time course of rapid transport along embryonic optic axons was carried out by reducing the efflux of labelled proteins from the eye by giving VLB intravitreally 2 h after the pulse. It revealed some peculiar features in the retino-tectal migration of glycoproteins and confirmed their progressive accumulation within terminals as previously described by radioautography. These results suggest that axonal transport of proteins during embryonic life undergoes changes in parallel with synaptic maturation. It may thus be considered as one of the factors controlling the genesis of neuronal networks.


A comparison of aldehyde fuchsin and alcian blue staining of neurosecretory material in Oncopeltus fasciatus. The dynamics of the "A" cells of the parsintercerebralis of Oncopeltus fasciatus over the first eight days of adult life was studied by microspectrophotometry of sections stained either with aldehyde fuchsin or alcian blue 8 GX. The data show that the two stains differ in their selectivity as they record different events in the history of the cells. A hypothesis is proposed that the aldehyde fuchsin is more sensitive to the presence of a "carrier" protein in the cell, whereas alcian blue 8 GX is more sensitive to the presence of the "active principle" in the cell.


Effects of barbiturates on ultrastructure and polymerization of microtubules in vitro. Barbiturates were examined for in vitro effects on ultrastructure of the frog sciatic system and polymerization of microtubules (MT) in a brain supernatant. Exposure for 5-17 h to 2.0 mM barbiturates caused a considerable loss of MT in ganglionic cell bodies and sciatic axons. This was mostly followed by a proliferation of 10 nm filaments. Under similar conditions treatment with 1 mM NaCN or 0.1 mM 2,4-DNP did not change the number or ultrastructure of MT and filaments. Eight barbiturates, varying in binding ratios to serum albumin and partition coefficients, were tested for effects on polymerization of MT using viscometry. Inhibitory effects were found which correlated with their reported ability to bind to albumin and brain fractions. Dimethylsulphoxide and ethanol were used as solvents for some of the barbiturates. These solvents at 1% had stabilizing effects on MT. The present results are discussed in relation to previous findings of inhibition of rapid axonal transport in vitro in the frog sciatic system by barbiturates.


Two active forms of RD-114 virus DNA polymerase in infected cells. Two forms of DNA polymerase are present in RD-114-infected human, dog, and mink cells, but are not detectable in uninfected cells. The two enzymes are indistinguishable catalytically and immunologically, but differ with respect to molecular weight and elution position from (dT)12-18-cellulose and phosphocellulose. The large enzyme (equivalent 95,000 daltons) is found in the infected cells, but not the virions produced by these cells. The virions contain only the smaller enzyme (equivalent 70,000 daltons). The larger form may represent a mammalian viral equivalent to the beta subunit of avian RNA tumor virus DNA polymerase.


The control of serum protein synthesis in hepatoma-fibroblast hybrids. Hybrids between mouse hepatoma cells (which secrete several serum proteins) and mouse or rat fibroblasts (which do not secrete these proteins) produce transferrin and the third component of complement (C3) like the parental hepatoma cells, while they do not secrete either albumin or alpha-fetoprotein (AFP). This lack of albumin and AFP secretion is probably due to a lack of synthesis, rather than to a simple defect in secretion. The cessation of albumin and AFP production is not dependent upon the parental fibroblast nor upon the selection conditions; it is best explained by a shut-off synthesis and could thus reflect the existence of a regulatory mechanism. This would imply a difference between the control of albumin and AFP synthesis and that of transferrin and C3 synthesis. On the other hand, in agreement with Peterson and Weiss (1972), hybrids between rat hepatoma cells and mouse fibroblasts continue to product rat albumin. This suggests that the mouse hepatoma cells differ from the rat hepatoma cells in the way they control albumin production.


[Scanning transmission electron microscopy of dendritic spines stained by the Golgi method]. Scanning transmission electron microscopy of the dendritic spines of multipolar neurons in the cat inferior Colliculus was achieved on Golgi semi-thin sections. The three basic types of dendritic spines (ST, MS, TH) were identified. Scanning transmission electron microscopy provides a reliable method for a three dimensional view of these structures at high resolution and consequently a more accurate appreciation of their size. In addition, it could prove very useful in the quantitative analysis of the dendritic spines.


[Metabolism and in vitro binding of several organochlorine and organophosphate pesticides to calf thymus DNA and rat liver microsomal proteins]. The organophosphorus insecticide ethyl-parathion was transformed by rat liver microsomes into metabolites which were bound to calf thymus DNA, in large amount. When the rats were treated by phenobarbital or 3-methylcholanthrene, the metabolite binding to DNA was increased two-fold. By contrast, in the same conditions, the organochlorine insecticides, aldrin, dieldrin and gamma hexachlorocyclohexane (lindane), did not yield metabolites able to bind to DNA and to proteins.


[Effect of injection of a phosphorylcholine-containing compound from Nippostrongylus brasiliensis on the infestation of the rat by this parasite]. Rats were injected intradermally with the purified phosphorycholine bearing component of the nematode Nippostrongylus brasiliensis. Eight days later they were experimentally infected with 1 X 10(3) infective larvae of the parasite. Compared to the non injected controls we showed a significant decrease in the number of adult worms able to be established in the small intestine. This protective effect seems to be due at least partly to a phenomenom being located between the skin and the lungs of the rats.


Report of the Inter-Society Commission for Heart Disease Resources. Optimal resources for cardiac surgery guidelines for program planning and evaluation. This updated and expanded planning guideline describes the optimal environment in which cardiac surgery can be performed effectively and safely and suggests criteria for evaluating existing facilities and for long-range hospital planning. The status of cardiac surgery is reviewed and measures are provided for assessing a hospital's ability to support a cardiac surgical service. Recommendations are given for appropriate case loads, administrative arrangements, and professional staff qualifications and relationships. The emerging role of the nurse is considered and requirements are enumerated for allied clinical supporting services. Special criteria are provided for pediatric cardiac surgical units and detailed specifications are given for the hospital's physical plant and equiptment including a protocol for checking the electrical safety of surgical suites. A data base is suggested for taking regular inventory of community cardiac surgical programs.


Fetal hypothyroidism complicating medical treatment of thyrotoxicosis in pregnancy. Two women with thyrotoxicosis were treated with antithyroid drugs during pregnancy. One women had inadvertently received a therapeutic dose of radioiodine at 21 weeks gestation and the other suffered from severe thyrotoxicosis with a serum LATS level of 1850%. In both patients, the serum triiodothyronine was maintained above 500 ng/dl by the concurrent oral administration of this hormone. Despite this precaution, cord serum thyrotrophin levels were markedly elevated and both infants showed clinical signs of hypothyroidism at birth. This experience indicates that triiodothyronine does not prevent fetal hypothyroidism when given to the mother in pharmacological amounts.


Association of fibrinogen and microfibrils with trophoblast basement membrane. Two non-collagenous placental antigens have been detected both in membranes isolated from terminal villi by sieving and sonication and in acetic acid extracts of the villi. One of the antigens is apparently fibrinogen based on immunologic and chemical data. The second, detectable with anti-membrane antiserum after absorption with fibrinogen remained with the fibrinogen-like antigen during isoelectric precipitation at pH 4.7, electrophoresis at pH 8.6 and chromatography on DEAE cellulose. Fractions of villi in which the two antigens occur comprise more than a third of the weight of the villi. The fibrinogen-related antigen was concentrated in placental basement membranes compared to kidney and lung basement membranes. A third antigen common to the membranes, to glomerular basement membrane and to alveolar basement membrane was also detected.


Immunochemical studies of organ and tumor lipids XXI. Sensitivity and specificity of the cytolipin F - sheep erythrocyte system. The sensitivity and specificity of the sheep erythrocyte - anti-sheep erythrocyte system to inhibition by pure cytolipin F has been studied with 5 antisera, in order to compare it with the rat erythrocyte-anti-rat lymphosarcoma system and its inhibition by pure cytolipin R. The cytolipin F - sheep erythrocyte system is much more sensitive than the cytolipin R - rat erythrocyte system, inhibition of hemolysis of 6 x 10(6) sheep cells being produced by 10 ng of cytolipin F (combined with a four-fold quantity of lecithin) compared with inhibition of hemolysis of 10(6) rat cells by 50 to 100 ng of cytolipin R (also combined with lecithin). Differences in sensitivity are attributed to the larger number of available cytolipin F determinants on sheep erythrocytes compared with cytolipin R determinants on rat erythrocytes. Studies of the auxiliary lipid enhancement of cytolipin F activity by galactocerebroside, lactosyl ceramide (cytolipin H), and lecithin are also reported.


Palliative esophageal intubation for carcinoma. A combined push-pull method. The use of prosthetic tubes for palliation of esophageal carcinoma can be carried out with safety using a combined push-pull method. The advantages of simultaneous endoscopy and laparotomy are illustrated in two cases. For safe durable palliation, emphasis must be placed upon seating the tube under direct vision as well as upon the firm fixation of the tube of the abdominal wall to prevent its subsequent migration.


Mechanisms of chromosome banding. VII. Interaction of methylene blue with DNA and chromatin. The binding of methylene blue to DNA and chromatin treated in various ways was examined by equilibrium dialysis. The maximum r value (moles of bound dye/mole of nucleotide) was 1.0 for DNA, 0.6 for unfixed chromatin, and 0.83 for chromatin fixed in methanol-acetic acid. When fixed chromatin was treated with saline-citrate at 60 degrees C for 3 hours, as used for G-banding chromosomes, the r value decreased from 0.83 to 0.55. When unfixed chromatin was treated as for R-banding the r values also dropped. Equilibrium dialysis indicated there was no disproportionate increase of dye binding as the concentration of DNA increased. -- These results, and others, suggest that some of the Giemsa negative regions of G- and R-banded chromosomes are due to the denaturation of non-histone proteins so that they more effectively cover the DNA and prevent side binding of the thiazin dyes.


[Clinical aspects of carcinoma of the liver (author's transl)]. The incidence of epi-cirrhotic liver carcinoma is apparently on the increase. Between 1969 and 1973 there were 84 cases of primary liver carcinoma and 467 of cirrhosis among 10211 autopsies. In 74 (16%) the carcinoma had developed in a cirrhotic liver. Clincally the diagnosis of carcinoma had been made in only 14 cases. The mean survival time after the diagnosis was seven weeks, after onset of the terminal symptoms 13 weeks. Typical clinical features were decompensated, hypertrophic, often coarsely granular, liver cirrhosis. The best diagnostic method was apparently laproscopy and determination of alpha 1-fetoprotein and cholestatic enzymes.


[Patho-physiological aspects in csf proteins (author's transl)]. Electrophoresis and immunoelectrophoresis of cerebrospinal fluid of 201 patients with inflammatory CNS diseases revealed that over a total protein value from 0 to greater than 1, 2 g/1 there was merely a decrease in pre-albumins and a difference in the gamma-globulins as a pointer to immune reactions. Comparison with the absolute values indicated an increase of all fractions in inflammatory and non inflammatory diseases caused by impairment of the blood-CSF barrier. Again there was evidence of immune reaction in the gamma-globulin range. Impairment in the blood-CSF barrier was demonstrated by an increased concentration of IgG, IgA and IgM. An immune reaction was revealed by higher IgG values.


Visual cortex responses to abrupt changes in the periodicity of rapidly intermittent light. Visually evoked potentials that are stimulated by period-jumps in a high-frequency modulated luminance have been detected and studied experimentally in monkeys. The visual responses are shown quantitatively by a Principal Components analysis to depend remarkably simply, in shape and in amplitude, on the period-jump stimulus. The response shape is nearly invariant, except that the early part of the response shows a distinctive (though slight) dependence on the sign of the period-jump. The response amplitude is approximately proportional to the magnitude of the period-jump of a given polarity. These features of the evoked potentials demonstrate an intimate association with a suprafusion transient phenomenon that has been observed psychophysically in man. The experimental behavior of the visual potentials also agrees well with theoretical predictions.


An EEG simulator-a means of objective clinical interpretation of EEG. In 1969 Zetterberg presented a method for describing the spectral properties of an EEG signal, starting from the assumption that the signal is essentially stationary during the analysis epoch. The method involves determination of the parameters for a model consisting of a lenear filter with the ability to produce a signal with the same spectral properties as the EEG signal. Zetterberg and Ahlin described in 1975 an analogue simulator based on this model theory. With this simulator it is possible to reproduce practically all types of stationary, as well as a number of non-stationary, EEG signals. We have used this simulator to demonstrate in an illustrative manner the relation between the properties of a signal in the spectral domain and in the time domain. We have also endeavoured to anser the question: if one starts from the frequency spectrum, what does the EEG signal corresponding to this spectrum look like? We also draw attention to the usefulness of the simulator in connection with training and as an instrument for testing computer and other systems of EEG analysis.


Genetic effects in the cortical auditory evoked potential: a twin study. An experiment was carried out on 40 pairs of adult male twins to investigate the extent of genetical determination in the cortical auditory evoked potential. Tonal stimuli of 1 sec duration, at an intensity of 95 dB and a frequency of 100 c/sec, were used. The inter-stimulus interval was 33 sec and the bipolar evoked potential was measured between the CZ and T3 scalp positions. The reliability of the seven latency and amplitude measures was also calculated and this was taken into account in the subsequent genetic analysis. The biometrical gentical approach, which gives maximal information particularly on small samples, was used to analyse the data. A strong genetic influence was found on all the amplitude scores. The environment made no significant contribution to these. For the latencies there was some evidence for a genetic effect; however, this was not very strong and can well be interpreted as being due to between family environmental effects.


Amygdala and masseteric reflex. I. Facilitation, inhibition and diphasic modifications of the reflex, induced by localized amygdaloid stimulation. The changes in amplitude of the monosynaptic masseteric reflex (MR), induced by stimulation of the amygdaloid area for the defence reaction (N. basalis, pars magnocellularis) and in other subdivisions of the amygdaloid complex, were studied in cats with spinal section maintained under Flaxedil. Simultaneously, the the effects of stimulation on the tonic activity of the masseteric nerve were observed. A maintained facilitation of the MR was elicited by stimulation  of the lateral nucleus, the parvocellular portion of the basal nucleus and the cortical nucleus, while the reflex was inhibited during stimulation of the medial-most portion of the  posterior amygdala. Diphasic changes of the MR amplitude (initial facilitation followed by delayed inhibition) were regularly observed when stimulating the magnocellular portion of the basal nucleus. These diphasic changes were closely correlated with the previously described diphasic resporatory and cardiac responses elicited from the same are (Bonvallet and Gary Bobo 1972). The initial facilitation probably corresponds to the "alerting" stage of the defence reaction and the delayed inhibition, associated with cortical, respiratory and cardiac activation, to the "defensive" stage of the reaction. Stimulation of the same area also provokes tonic or rhythmical discharges of the masseteric motoneurons which frequently occur during the delayed inhibition of the MR. The main efferent pathway mediating these motor effects is probably the ansa lenticularis.


Amygdala and masseteric reflex. II Mechanism of the diphasic modifications of the reflex elicited from the "defence reaction area". Role of the spinal trigeminal nucleus (pars oralis). It has been demonstrated (Gary Bobo and Bonvallet 1975) that long-lasting stimulation of the "amygdaloid area for the defence reaction" (basal nucleus, pars magnocellularis) elicits, after an initial facilitation, a delayed inhibition of the monosynaptic masseteric reflex (MR), while stimulation of the amygdalofugal fibers running in the ansa lenticularis provokes an immediate inhibition of the reflex. In present study, the structure which mediates these inhibitions has been identified. Using combined techniques of limited transections, localized coagulations and localized stimulation and recording, it has been demonstrated that these ingibitions are mediated by the rostral portion of the spinal trigeminal nucleus, the subnucleus oralis (NO). After localized coagulation of this nucleus, or after lesions which interrupt slectively the connections between the NO and the masticatory nucleus, long-lasting stimulation of the basal nucleus elicits only well maintained facilitation of the MR. Hence, the delayed decrease in amplitude of the reflex, observed during stimulation of the basal nucleus in the preparations with intact brain, cannot be explained by the reversal of an initial facilitatory influence to an ingibitory one. The one ingibition of the reflex is due to the superimposition, on a background of sustained facilitation of the masseteric motoneurons, of the inhibitory influence exerted on the monosynaptic masseteric circuit by a trigeminal sensory nucleus, itself activated by delayed discharges of the basal nucleus. A tentative representation of the dual control exerted on the masseteric activity by the basal nucleus is given in Fig. 9. The functional implications of this dual control during the "defence reaction" are briefly discussed.


Differential effect of task relevance on early and late components of cortical and subcortical somatic evoked potentials in man. The effect of "task relevance" on early and late components of cortical and subcortical somatic evoked potentials (SEPs) was studied in a group of Parkinsonian patients operated on under local anesthesia for treatment of prominent unilateral tremor. 1. SEPs produced by median nerve stimulation were found at contralateral cortical (ss), thalamic (vcpci, vcai), lemniscal (Lm), postilemniscal (PoLm), prelemniscal (Raprl) and reticular (Ttc) regions. No SEPs were found in other contiguous thalamic (M,Pf, ce) and subthalamic (Q) regions. 2. Subcortical early SEP components consisted of two monophasic positive potentials distributed within a circumscribed thalamo-lemniscal region where electrical stimulation elicited consistent sensory responses circumscribed to contralateral hand and face. In contrast, subcortical late SEP components consisted of monophasic or polyphasic, positive or negative potentials distributed in a widespread, thalamic, lemniscal, prelemniscal and reticular region where elecrical stimulation elecited sensory or motor responses of various types. Subcortical early and late SEP components appeared together in lemniscal, thalamic and cortical regions but they wers separated at postlemniscal (only early) and prelemniscal and reticular ones (only late). 3. Significant amplitude changes in cortical and subcortical late SEP componets were found concomitant to variations in "task relevance": they decreased when patients shifted from novelty to habituation, they increased when patients shifted from habituation to attention and they decreased when patients shifted from attention to distraction. In contrast, no significant ampiltude changes in cortical and subcortical early components were found when patients shifted through these various "task relevance" conditions.


Local hypothalamic blood flow during sleep. There are several hypotheses to explain the observed rise in hypothalamic temperature during REM sleep. One of theories suggests that the increase in temperature is due to an increase in local blood flow. In order to test this hypothesis local hypothalamic blood flow (HBF) was measured during wakefulness, HBF increased by 63% and 25% during REM and non-REM sleep respectively. It is possible that the increase of HBF during REM sleep is due to the activation of the recently described noradrenergic pathways from the brain stem to the cortex.


Frequency and time domain characteristics of single muscle fibre action potentials. Action potentials of single muscle fibers (brachial biceps) were recorded with a multi-electrode, ensuring a minimum fibre-electrode recording distance. Propagation velocity was measured in 15 fibres and the power spectral density of the action potentials was computed through a FFT algorithm. Linear correlation was found between action potential amplitude (peak to peak) and propagation velocity, as well as between maximum-minimum amplitude time (defined as the time interval between the positive and negative peaks) and propagation velocity. Assuming al linear dependence between propagation velocity and muscle fibre diameter, a linear relation between the fibre thickness and the extra-cellular action potential was derived from the action potential amplitude/propagation velocity curve. The action potential power spectrum had a band pass form, with peak magnitude of 1.61+/-0.03 kc/sec, and -3 dB points at 0.98+/-0.19 kc/sec and at 2.41+/-0.53 kc/sec. A linear relation was found between the spectral peak magnitude and propagation velocity, and between the ban width and the propagation velocity, as predicted by a mathematical model describing the power spectrum of single fibre action potentials.


Quantitative assessment of the electroencephalogram in renal disease. EEGs wre recorded from renal patients to determine if there are quantifiable characteristic changes in the EEG was quantified by calculating the percentage of spectral power in the bandwidth 3-7 c/sec referrred to a frequency range of 3-13 c/sec and by computing the mean frequency of the dominant rhythm in the EEG. Blood urea nitrogen and creatinine concentrations, as well as a self-assessment of the patient's clinical condition, were recorded. The general finding of this research is that EEG slowing, as evaluated by power spectral techniques, is correlated with uremia-associated variables. 1. In a non-dialyzed patient population with renal failure, slowing in the EEG was found to be directly corelated with increased creatinine concentrations. 2. Quantitative measures of slow wave activity computed using power spectral techniques were found to be highly corelated with an estimate of slowing made by an electroencephalographer. 3. Compared with undialyzed azotemic patients, malignant hypertensive patients with comparable serum creatinine concentrations typically displayed increased slow wave activity, while slowing was generally reduced in the dialyzed patient population. 4. A series of EEGs recorded from one patient during the first three dialyses of her life revealed that slow wave activity decreased during each successive dialysis. In another patient, all quantified EEG values recorded prior to renal transplantation significantly improved after transplantation...


Startle responses recorded in the leg of man. The EMG pattern of startle reactions in the leg was studied in man using a 100 msec, 1 kc/sec squre wave tone burst of 114 dB as the auitory stimulus. At rest, 74 responses were recorded in the ankle flexor tibialis anterior (TA) and 23 in the ankle extensor gastrocnemius (G) with a significantly different mean latency of 151 and 123 msecrespectively. During tonic extension or flexion of the ankle, the EMG response in TA or G consisted in a burst of activity superimposed on the voluntary EMG at a latency of approximately 150 msec after the onset of the tone burst and followed, at 200 msec, by a period of EMG silence lasting close to 100 msec. Similar periods of silence also occurred as the sole EMG modulation in response to sound. Although predominant in flexors, startle reactions are significant in extensors during or after tonic ankle extension suggesting that audiospinal influences may be channelled to one or the other group depending on their functional state. The silent period of EMG following an excitatory startle or appearing on its own suggests an important inhibitory component in audiospinal mechanisms particularly related to startle since it had not been disclosed in a previous study of the modulation of the H-reflex by non-startling auditory stimulus. From the distribution and EMG pattern of startle, it is inferred that audiospinal influences may be meaningfully integrated through the reticulospinal system in sound guided behaviour.


The adaptation of cochlear and brainstem auditory evoked potentials in humans. The adaptation of the responses from the cochlear nerve and the auditory brainstem nuclei was studied using a burst of four clicks as the stimulus. Nine experimental conditions were obtained from three stimulus levels (60, 70 and 80 dB SL) and from three intervals between click in the burst (15, 24 and 32.5 msec). Six subjects were each tested three times for each of the experimental conditions and the techniques used and results obtained are summarized. Various models of the adaptation of the brainstem responses are proposed which predict different results. The results for the adaptation of the N1 response are basically in agreement with previous work, and the more central brainstem responses showed less adaptation than the peripheral responses. These findings may be explained by postulating different adaptation mechanisms for the peripheral and central responses. The 60dB stimulus level condition gave rise to more adaptation than the 80dB level, and this is in agreement with studies of low and high threshold reception systems reported elsewhere.


A dual marking technique for microelectrode tracks and localization recording sites. This paper describes techniques for marking both microelectrode tracks and exact recording loci using a combination of fast green dye and horseradish peroxidase (HRP). The procedure involves coating the exterior of HRP filled microelectrodes with fast green dye in order to identify electrode tracks, and ejecting HRP from the electrode to mark recording loci. Rapid, multiple marks can be made with this technique without harming the recording capabilities of the micropipette.


A technique for on-line classification of evoked potentials into two groups according to subjective interpretation of the stimulus. A technique is described for on-line classification of evoked potentials (EPs) into distinct groups for separate averaging according to a criterion available after the beginning of the EPs. It is suitable for analysing psychophysiological correlations (relationship between EPs and subjective perception). The system consists of a slightly modified NTA 512 B (Hungarian) average response computer (however, any CAT-computer-like device can be used in a similar way) and of an additional device. It can be used in all cases when the number of subjective judgements according to which EPs have to be classified can be limited to two (or three, when, by slightly changing the system, all quarters of memory are used).


A digital instantaneous impulse rate meter for neural activity. An instantaneous impulse rate meter is described, which can be used fro the precise instantaneous conversion of neural or heart activity trains in real time. The device is designed in digital technology to meet various requirements which are important for physiological reserach laboratories.


[A case of reading epilepsy: effects of attention and reading tests on EEG and "H" reflex]. The effects of attention and reading tests on the EEG and the H reflex were studied in a patient affected by primary reading epilepsy. During the attention tests, EEG was normal while H reflex was facilitated. On the contrary, during reading tests, EEG showed typical epileptic seizures while at the same time, the H reflex was depressed.


Soleus H reflex and afferent discharges from antagonist muscles. An electrophysiological method of evaluation. The influence of electric activation of the peroneal nerve (i.e., of anterior leg muscles) on the ipsilateral soleus H response was studied in normal, parkinsonian, hemiplegic and spastic spinal paraplegic subjects. The anterior muscles were held in a forced extended position to obtain their maximal stretch during isometric contraction. The stimulus evoking soleus H reflex was delivered after a conditioning stimulus to the peroneal nerve at different intervals, between 10 and 500 msec. The excitability curve of the soleus H reflex obtained in this way showed a rather constant pattern in normal subjects, which significantly differed from that of the parkinsonians. "Pyramidal" patients showed a pattern not significantly disimilar from normals. Parkinsonians were characterized by clear facilitation of the soleus H response for intervals (about 100-150 msce) corresponding to the maximum tension increase on the anterior muscles.


[Diurnal rhythms of thyroid hormones in rooster chicks (Gallus domesticus). 2. Studies on the protein-bound radioiodine and the "free" hormone portion]. In four week old cockerels the plasma was investigated for cyclic changes in total radioactivity, PB131I, PI131I, and "free" thyroid hormones 24 hours after injection of Na131I. Maxima in total radioactivity were observed at 3.00 am and 9.00 pm. They were significant different from the minima at noon (2P less than 0.005). At the same points of the day maxima, resp. minima were found in the PB131I and the "free" hormones. The "free" hormones expressed in per cent of total hormones showed low values at 6.00 am, 3.00 pm, and 9.00 pm. The diurnal changes in haematocrit and albumin concentration were not responsible for the variation of PB131I. In order to eleminate effects of isotope dilution by ingested iodine the PB131I was expressed in per cent of total radioactivity of plasma (= conversion rate) or resp. in per cent of thyroidal radioiodine uptake. The obtained values showed maxima at 6.00 am and 9.00 pm. From that we conclude again on a stimulation of the thyroid at these times.


Inhibition of thyroid function and thyrotrophic activity following the administration of methallibure (ICI-33 828). 1. Goitrogenic action of methallibure (ICI-33828) has been studied in mice gerbil and hedgehog using thyroid weight and histological structure as an index. Liquefaction and vacuolation of thyroid follicles were most prevelant after methallibure administration. 2. The I131 content of the thyroid gland was significantly higher in the methallibure treated groups than in the controls. This denotes a decrease in the rate of discharge of thyroid hormone. 3. Protein bound radioiodine (Pb I131) was low after methallibure administration. 4. Methallibure administration brings about hypertrophy of pituitary thyrotrophs which is also reflected in the increased basophilic cell percentage in gerbil (control: 15.5 percent; methallibure: 22.8 percent). 5. It is concluded that methallibure acts on thyroid function both by a direct effect on the gland as well as by influencing pituitary thyrotrophic activity in enhancing I131 uptake.


Mouse immunoglobulin genes: studies on the reiteration frequency of light-chain genes by hybridisation procedures. The partially purified immunoglobulin light chain messenger RNA fraction from P3K (MOPC 21) mouse myeloma tissue-culture cells has been employed in hybridisation studies. Fragments of the messenger RNA were generated by alkali hydrolysis. 6-S fragments not containing poly(A) showed the characteristic biphasic hybridisation profile seen with the intact RNA fraction. 12-S and 6-S poly(A)-containing fragments, however, showed single transitions lacking the rapidly hybridising component. Complementary DNA copies of the intact messenger RNA fraction were prepared with RNA-dependent DNA polymerase and the DNA populations fractionated on acrylamide gels. Hybridisation experiments with complementary DNA fractions up to 800 bases in length showed annealing to single (or a few) genes. A rapidly hybridising component (about 200 copies) appears in the cDNA fraction containing the largest transcripts. We conclude that the kappa constant region gene and the MOPC 21 variable region gene are present as one or a few copies in the haploid genome and that the rapidly hybridising component is not due to variable region genes.


Distinct ribonuclease H activities in calf thymus. Three enzymes with ribonuclease H activity are present in calf thymus. They have been separated on the basis of chromatographic behaviour and molecular weight. They are further distinguished from one another by their ionic requirements and sensitivity to the -SH reagent N-ethylmaleimide. Two of these enzymes are classified as ribonuclease H, the third is obtained in a fraction which possesses ribonuclease H activity but also degrades double-stranded RNA and poly(rA). No association between any of the enzymes and cellular DNA polymerases could be demonstrated.