Release of beta-thromboglobulin from human platelets by therapeutic intensities of ultrasound. The effects of therapeutic intensities of ultrasound on human platelets in whole blood were investigated by monitoring the release of the platelet specific protein beta-thromboglobulin (beta-TG). More beta-TG was released as the intensity of the ultrasound was increased and also as the driving frequency was decreased from 3.0 to 0.75 MHz. Some beta-TG was released at spatially-averaged intensities as low as 0.6 W/cm2 at 0.75 MHz, a value significantly lower than that observed for the onset of aggregation of platelet rich plasma (obtained from the same volunteer) in the same exposure system. Liberation of beta-TG by ultrasound was diminished but not abolished in the presence of inhibitors which rendered the platelets functionally inert. Our data suggests that beta-TG is liberated in two ways, firstly as a result of platelet disruption by cavitation, and subsequently by potent aggregating agents, liberated in parallel with beta-TG, inducing the physiological release reaction in adjacent platelets. The low therapeutic intensities and short exposure times (30 s or less) necessary to liberate beta-TG from normal human platelets in vitro, suggests that patients with abnormally sensitive platelets and/or 'hypercoagulable state' could be at risk if subjected to high therapeutic intensities of ultrasound.


Fundus signs in temporal arteritis.

Fundus signs in temporal arteritis. A patient with temporal arteritis developed a variety of ischaemic lesions in the eyes. Infarction of the inner retina and optic nerve head was delineated on presentation by white swelling in the retinal nerve fibre layer. The role of interrupted axoplasmic transport in the production of this sign is discussed. Outer retinal infarction was also noted on presentation and subsequently gave rise to striking pigmented scars.


Transition-metal binding site of bleomycin A2. A carbon-13 nuclear magnetic resonance study of the zinc(II) and copper(II) derivatives. The 13C NMR spectra at 25.2 MHz of the Zn(II) and Cu(II) complexes of the antitumor antibiotic bleomycin A2 are discussed. Complexation of the drug to Zn(II) causes 38 of the 52 resonance lines of bleomycin A2 to shift to new positions. All but ten of these shifted lines have been assigned in the Zn(II) bleomycin complex. Although the specific donor sites of the drug cannot be identified from the 13C NMR data, the analysis clearly shows that the pyrimidine-imidazole portion of the molecule is affected by chelation. This finding is in agreement with the previously reported metal-binding site of the antibiotic. The analysis also shows that carbon atoms which have large through-bond distances from the binding site can experience substantial chemical-shift changes upon metal binding. Complexation of the drug to Cu(II) eliminates 23 resonances from the spectrum of the molecule. All of these resonances emanate from carbon atoms which are located in the pyrimidine-imidazole portion of the drug.


Number of water molecules coupled to the transport of sodium, potassium and hydrogen ions via gramicidin, nonactin or valinomycin. The number of water molecules (n) coupled to the transport of cations across lipid membranes was determined in two different wats: directly from the electro-osmotic volume flux per ion, and by the use of Onsager's relation, from the open circuit streaming potential produced by an osmotic pressure difference. The results of the two approaches were in general agreement. Monoolein membranes were formed on the ends of polyethylene or Teflon tubing connected to a microliter syringe and the volume change necessary to keep the membrane at a fixed position was measured. It was necessary to make corrections for unstirred layer effects. The results for gramicidin were: n approximately 12 for 0.15 M KCl and NaCl, n approximately 6 for 3.0 M KCl and NaCl, and n approximately 0 for 0.01 M HCl. For nonactin, n approximately 4 for both 0.15 and 3.0 M KCl and NaCl. Valinomycin (for 0.15 M KCl) behaved like nonactin. It is shown that for a channel mechanism, in general, n is less than or equal to the number of water molecules in a channel that does not contain any cations. Thus, the n of 12 for the 0.15 M salts implies that the gramicidin channel can hold at least 12 water molecules. This places an important constraint on models of the channel structure. The n of 0 for HCl is consistent with a process in which protons jump along a continuous row of water molecules. The decrease of n with the 3.0 M salts may indicate that the channel becomes multiply occupied at high salt concentrations. The n of 4 for nonactin and valinomycin means that at least four water molecules are associated with the carrier . cation complex, probably in the interstices between the complex and the disordered lipid.


The effect of gramicidin A on the temperature dependence of water permeation through liposomal membranes prepared from phosphatidylcholines with different chains lengths. The permeation of water through liposomal membranes composed of various saturated phosphatidylcholine plus gramicidin A was studied as a function of temperature. 1. The presence of gramicidin in the liposomal bilayers caused an increase in water permeability. Below the phase transition temperature this effect could be measured quite clearly in all the systems we tested, but the extent of the increase was largely dependent on the length of the hydrocarbon chains. 2. Increasing amounts of gramicidin caused a gradual disappearance of the abrupt change in the rate of water permeation near the gel-liquid crystalline phase transition temperature of dipalmitoyl phosphatidylcholine liposomes. Differential scanning calorimetry analysis of the system containing these relatively small amounts of gramicidin still showed a clear transition from the liquid crystalline to the gel state with only a slight reduction in the enthalpy change. 3. In liposomes composed of dimyristoyl, dipalmitoyl and saturated egg phosphatidylcholine there was a concomitant decrease in the activation energy of water permeation in the presence of gramicidin below and above the phase transition temperature. The activation energy for water permeation through longer chained distearoyl phosphatidylcholine liposomal bilayers was the same with or without gramicidin in the bilayer. 4. It is concluded that the ability of gramicidin to form conducting channels in a gel state bilayer depends on the thickness of the paraffin core.


Heterogeneity of human alpha-fetoprotein as revealed by isoelectric focusing in urea-containing gels. The heterogeneity of human alpha-fetoprotein has been studied by analytical isoelectric focusing in polyacrylamide gel slabs in the presence of 8 M urea. Six major isoelectric variants could be identified over a pH range of 6.0--6.2. Verification of their identity was achieved by crossed immunoelectrophoresis into agarose gel containing monospecific antiserum to human alpha-fetoprotein. Complete desialylation of the protein did not abolish the heterogeneity; a complex pattern of major alpha-fetoprotein bands persisted over a more alkaline pH range. We have been able to correlate the pattern of alpha-fetoprotein heterogeneity seen following extended agarose gel electrophoresis with that obtained during isoelectric focusing in the presence of urea. The quantity of certain alpha-fetoprotein charge isomers in various alpha-fetoprotein isolates may be important in considering certain biological functions of this protein.


Synthesis of a fully active snake venom cardiotoxin by fragment condensation on solid polymer. A polypeptide cardiotoxin containing 60 amino acids with 4 disulfide bonds has been synthesized by the "fragment solid-phase" method. The identity of the synthetic product with native cardiotoxin was established by chromatography on Sephadex G-50, carboxymethyl-cellulose column chromatography, thin layer chromatography, disc gel electrophoresis, amino acid analysis, end group analysis, peptide mapping, circular dichroism spectra, and four biological tests.


[Study of individual Pacinian corpuscle mechanoreceptors following application of colchicine to a nerve]. Colchicine application to the cat caudal mesenteric nerve containing sensory fibers for single mechanoreceptors (Pacinian corpuscles) causes degeneration of the axis cast of the nerve endings. Ultrastructural changes in the receptors showed no difference from the axonal degeneration after the nerve section but the rate of degeneration was considerably slower. Ultrastructural, electrophysiological, and biochemical changes occurring in the Pacinian corpuscles were not the result of direct action of colchicine, but appeared to be realized through the nerve by the axoplasmic transport block. It is suggested that the receptor's structure is under the sensory neuron neurotrophic control.


[Inhibition of non-cytotoxic histamine liberation from isolated rat mast cells by antihistaminic preparations]. Phenothiazines (chlorpromazine and promethazine) and antihistaminic quinuclidine derivatives [phencarol, quinuclidyl-3-di-(o-tolyl) carbinol, hydrochloride quinuclidyl-3-di-(o-methoxyphenyl) carbinol--HQMC] at concentrations preceding the histamine-releasing ones inhibited the compound 48/80-induced histamine release from the isolated rat mast cells. HQMC inhibited histamine release induced by selective liberators (compound 48/80, MCD-peptide, specific antigen), but potentiated histamine release induced by nonselective liberators (chlorpromazine, tryton X-100). The inhibition by HQMC of histamine release induced by compound 48/80 increased during 1 min and was reversible. The inhibitory effect of all the compounds tested was partially counteracted by glucose.


Gammaglobulin treatment and anti-IgA antibodies in IgA-deficient patients. Antibodies to IgA may cause severe anaphylactic reactions during blood transfusions. Tests for anti-IgA antibodies were carried out on six patients with IgA deficiency (five of whom also had hypogammaglobulinaemia) who had received continuous gammaglobullin treatment for chronic or recurrent infections for three to eight years. Three patients had minute amounts of IgA, and three had none (less than 0.01 microgram/ml). Only one patient had anti-IgA. Her antibody titre did not change during treatment. No patient had any untoward effects of treatment, which relieved the symptoms of infection in every case. IgA determinations should be performed by more accurate methods than radial immunodiffusion when evaluating the risks of giving gammaglobulin to patients with hypogammaglobulinaemia and IgA deficiency. Probably the stimulus provided by intramuscular gammaglobulin in such patients is insufficient for the formation of anti-IgA antibody.


Detection of a new antibody system reacting with Dane particles in hepatitis B virus infection. Antibodies in the serum reacting with antigens on the surface of radiolabelled Dane particles distinct from hepatitis B surface and core antigens (HBsAg and HBcAg) were detected, using a double antibody precipitation assay, in 12 out of 15 patients early in the course of acute type B hepatitis and at the time of disappearance of circulating Dane particles. No such antibody activity was found in 15 of the 16 patients with HBsAg-positive chronic active hepatitis, 13 of whom had complete Dane particles in the serum. In a group of 16 asymptomatic HBsAg carriers (without Dane particles in serum) antibody activity was shown in nine. This demonstration of antibodies precipitating Dane particles may be relevant to the clearance of circulating hepatitis B virions and the termination of infection in acute type B hepatitis. Their absence in all but one of the cases of chronic active hepatitis might explain why the virus infection persists in this group of patients.


Regeneration of motor axons in the rat sciatic nerve studied by labeling with axonally transported radioactive proteins. Labeling regenerating axons with axonally transported radioactive proteins provides information about the location of the entire range of axons from the fastest growing ones to those which are trapped in the scar. We have used this technique to study the regeneration of motor axons in the rat sciatic nerve after a crush lesion. From 2 to 14 days after the crush the lumbar spinal cord was exposed by laminectomy and multiple injections of [3H]proline were made stereotactically in the ventral horn. Twenty-four hours later the nerves were removed and the distribution of radioactivity along the nerve was measured by liquid scintillation counting. There was a peak of radioactivity in the regenerating axons distal to the crush due to an accumulation of label in the tips of these axons. After a delay of 3.2 +/- 0.2 (S.E.) days, this peak advanced down the nerve at a rate of 3.0 +/- 0.1 (S.E.) mm/day. The leading edge of this peak, which marks the location of the endings of the most rapidly growing labeled fibers, moved down the nerve at a rate of 4.4 +/- 0.2 mm/day after a delay of 2.1 +/- 0.2 days; this is the same time course as that of the most rapidly regenerating sensory axons in the rat sciatic nerve, measured by the pinch test. Another peak of radioactivity at the crush site, presumed to represent the ends of unregenerated axons or misdirected sprouts, declined rapidly during the first week, and more slowly thereafter.


Effect of glucosamine on virus production and antigen expression in avian sarcoma virus-transformed cells. Treatment by glucosamine of avian sarcoma virus-transformed chicken embryo fibroblast (CEF) cells completely inhibited the formation of progeny-transforming virus particles. Such cells, however, could continue to synthesize non-infectious physical particles containing both viral RNA and the enzyme RNA-dependent DNA polymerase if glucosamine exposure was performed in the presence of glucose. Glucosamine treatment was found to affect antigenic expression in transformed CEF as measured by an indirect immunofluorescence test. Inhibition to a far lesser extent was observed when a lymphocyte stimulation assay for the detection of cell-mediated immunity was used in this system.


The immunochemical complexity of CEA: a golden dream or molecular nightmare? Carcinoembryonic antigen is only a tumor assoicated antigen and is less clinically useful than the original report suggested. However, the CEA assays in clinical use utilize reagents operationally defined by old criteria. There is now abundant evidence that "CEA" consists of a heterogenous family of related glycoproteins with shared, as well as distinctive, antigenic determinants. Antigenic differences can be demonstrated between some cancer sera "CEA" and various operationally defined tissue "CEA" preparations. Further definition of the biochemical and antigenic characteristics of serum and tumor tissue "CEA" is required in order to determine whether a more tumor specific antigenic determinant can be identified.


Isolation of HLA and tumor antigens by means of affinity chromatography employing anti-beta2-microglobulin (beta2m) antiserum. A method for the isolation of HLA antigen molecules from normal and cancerous solid human tissue is described. The method employs anti-beta2-microglobulin (beta2m) antiserum coupled to Sepharose beads as an immunosorbent affinity medium. The anti-beta2m affinity chromatography procedure greatly purifies and selectively enriches HLA and any material that copurifies by affinity, with beta2m and/or HLA molecules. The HLA isolated by this purification procedure was used to immunize rabbits. The antisera obtained were absorbed on beta2m to remove all anti-beta2m antibody activity. The use of such anti-HLA antisera in radioimmunoassays, immunoprecipitation studies, and F(ab')2 blocking experiments demonstrated that these antisera are directed against a common HLA determinant present on the heavy (alloantigen-bearing) chain of all HLA molecules. The use of an identical procedure employing human tumor tissues has resulted in the isolation of HLA-like or HLA-associated tumor-specific antigens as demonstrated by the leukocyte adherence inhibition (LAI) assay.


Ovarian tumor antigens.

Ovarian tumor antigens. Evidence has been reported for at least two common tumor-associated antigens, or antigenic determinants, in human cystadenocarcinomas of the ovary that are apparently absent in tissues of normal reproductive organs. These antigenic determinants are immunologically distinct from carcinoembryonic antigen, alpha-fetoprotein, ferritins and histocompatibility antigens. One of these two ovarian cystadenocarcinoma-associated antigens (OCAA) is not detectable in any ovarian carcinomas except serous or mucinous types, other gynecologic or nongynecologic malignancies thus far tested, while the second antigen is present in about 90% of all gynecologic tumors and occasionally in breast and colon tumors. OCAA has been purified and partially characterized. It is a high molecular weight glycoprotein which carries the unique ovarian tumor-specific antigenic determinant along with some normal cross-reacting determinants. High levels of this glycoprotein antigen have been detected in the sera of ovarian cancer patients with advanced disease by the radioimmunoassay inhibition technique. The serial determination of circulating OCAA appeared to correlate with tumor volume as well as the clinical status of the patients.


Further characterization of CSAp, an antigen associated with gastrointestinal and ovarian tumors. Colon-specific antigen-p, or CSAp, was originally extracted from GW-39 tumors, which are human colonic carcinomas serially transplanted in golden hamsters, and antibodies to CSAp have been produced in the same animal hosts. By means of immunodiffusion and a hemagglutination-inhibition assay, CSAp has been found to be restricted to adult and fetal small intestine, neoplastic gastric and colonic tissues, inflamed colon, and cystic mucinous tumors of the ovary. CSAp was shown to be distinct from blood group antigens, including Lea and Leb blood group substances, liver ferritin, AFP, CEA, CSA, CMA, ZGM, and BOFA, and to have the electrophoretic mobility of an alpha2-globulin. Gel filtration studies indicated that CSAp in GW-39 tumor, primary human colonic carcinoma, and ovarian cancer mucinous cyst fluid had a peak molecular size range of 70,000--110,000. Quantitation of CSAp in 214 tissue specimens by the hemagglutination-inhibition assay revealed a progressive increase in fetal, inflamed, and neoplastic intestine, such that CSAp in colonic tumors was increased over normal colon tissue. Thus, CSAp appears to be an organ-specific antigen showing increased levels in some gastrointestinal and ovarian neoplasms, as well as in specimens with colitis.


Combination chemotherapy of head and neck cancer. A total of 77 patients with cancer of the head and neck area were treated with five different drug combination regimens. Five of the 77 patients had lymphoepithelioma; four had adenocystic carcinoma, and 68 had squamous-cell carcinoma of the head and neck (16 from the skin). Of these 77 patients, 16 had no previous treatment, five had surgery, 11 had radiotherapy, and 45 had surgery and radiotherapy. The first regimen consisted of a four-day Bleomycin infusion followed after a 24-hour rest, by cyclophosphamide (Cytoxan), Vincristine (Oncovin), methotrexate and 5-Fluorouracil (5-FU) (B-COMF). The next three regimens consisted of a four-day Bleomycin course, followed by either Cytoxan and methotrexate (B-CM), Cytoxan and 5-FU (B-CF) or Methotrexate and 5-FU (B-MF). The fifth regimen consisted of Bleomycin concomitant with Cytoxan, Methotrexate, and 5-FU (B-CMF). Of the 49 patients receiving B-COMF and B-CMF, 12 showed a complete response and 12 a partial response. Among the 28 patients receiving Bleomycin, followed by any one of the two drug regimens, only six showed a partial response. The severity of the thrombocytopenia, number of drugs, lymphoepithelioma histology and performance status of the patient influenced the rate of response. Drug toxicity consisted mostly in myelosuppression. The B-CMF combination is highly effective and can be used as an adjuvant to surgery and/or radiotherapy.


Diffuse pulmonary malignant mesothelioma: response to doxorubicin and 5-azacytidine. A 46-year-old patient who developed a right pleural mesothelioma 18 years after asbestos exposure was found to have diffuse reticular nodular infiltrates in both lungs. Lung biopsy by fiberoptic bronchoscopy revealed alveolar and interstitial spread of the neoplastic cells. A dramatic clinical and radiologic response occurred after treatment with a combination of doxorubicin (Adriamycin) and 5-Azacytidine.


Combination chemotherapy against B16 melanoma: bleomycin/vinblastine, bleomycin/cis-diamminedichloroplatinum, 5-fluorouracil/BCNU and 5-fluorouracil/methyl-CCNU. Four antitumor drug combinations which are currently in clinical use were evaluated experimentally using the murine B16 melanoma model. Bleomycin plus vinblastine produced an increase in life span over either of the two agents alone against both intraperitoneal and subcutaneous B16. This combination also resulted in a large number of long-term survivors. Bleomycin plus cis-platinum produced slight enhancement against subcutaneous B16, but showed no advantage against intraperitoneal B16. The combination of 5-fluorouracil plus methyl-CCNU significantly increased survival time against the intraperitoneal tumor, and produced long-term survivors as well. The combination of 5-fluorouracil plus BCNU was not more effective than BCNU or 5-fluorouracil alone. These data were compared with the degree of success reported from the clinics against a variety of solid human neoplasms.


Immune response of prostatic cancer patients to cytomegalovirus-infected and -transformed human cells. The indirect immunofluorescent test was used to determine the prevalence of humoral immunity to cytomegalovirus (CMV)-induced antigens in prostatic cancer patients as compared to age-matched controls. Significantly more prostatic cancer patients demonstrated high CMV-antibody titers than did the benign prostatic hyperplasia and nonurogenital cancer groups; however, no significant difference in reactivity was found between paients with prostatic cancer and transitional cell carcinoma of the urinary bladder. When screened against CMV-transformed human cell lines, the reactivity of the sera followed the rate of expression of CMV-related antigens of cell lines used in these tests.


Variables for predicting serious toxicity (vinblastine dose, performance status, and prior therapeutic experience): chemotherapy for metastatic testicular cancer with cis-dichlorodiammineplatinum(II), vinblastine, and bleomycin. Combination chemotherapy utilizing high-dose vinblastine, bleomycin, and cisdichlorodiammineplatinum(II) (CDDP) is effective treatment for metastatic testicular cancer. Unfortunately, it is frequently associated with serious toxicity. In a series of 14 patients receiving 65 treatment cycles, several variables were examined as putative risk factors for prediction of serious toxicity. These included drug dose normalized to body weight or surface area, interval since previous cycle, prior therapeutic experience with either radiation therapy or other cytotoxic chemotherapy, and Karnofsky performance status. The strongest determinant of serious toxicity was the vinblastine dose calculated according to body weight. The next most influential prognostic variables were the performance status and a history of previous treatment with either radiation therapy or chemotherapy. Serious toxicity may be anticipated at a frequency of 40% when vinblastine is administered at a total dose of 0.36 mg/kg with bleomycin and CDDP. In our group of patients, the nephrotoxicity of CDDP appeared to be cumulative despite intensive diuresis at the time of administration. Pulmonary toxicity was not observed. Modest reductions in vinblastine dose, especially in patients with poor performance status or a history of previous radiation or other chemotherapy, will substantially lower the frequency of serious toxicity.


Appraisal of skin tests with food extracts for diagnosis of food hypersensitivity. Seventy-six children aged 5 months to 15 years who exhibited a net weal of 3.0 mm or greater to a puncture skin test with one or more of fourteen foods were subjected to double-blind food challenge. Confirmed reactions to double-blind food challenge were found to occur only with peanut, milk, egg and soybean. Puncture skin tests with 1:20 w/v concentration of food extracts identified all subjects who exhibited an adverse reaction during the double-blind food challenge. Performance of intradermal skin tests did not identify any additional subjects who reacted clinically to double-blind food challenge.


The application of photobleaching to sequential staining of antigen-antibody precipitates in gels for prostatic acid phosphatase and protein. A modified procedure is described for the sequential staining of agar precipitates of seminal plasma and antiseminal plasma serum for acid phosphatase and total protein. The procedure employs Gomori's glycerophosphate-lead sulphide method for acid phosphatase, ultraviolet light for photobleaching and amidoschwartz for total protein staining. In a single agar diffusion plate, a minimum of 7 protein bands was observed, 3 of which contained acid phosphatase activity.


A simple and reliable method for the purification of human alphafetoprotein (AFP) from amniotic fluid and fetal livers. Highly purified human alphafetoprotein has been isolated from amniotic fluid and fetal livers by a combination of salting-out, and gel filtration, ion exchange, and concanavalin-A affinity chromatography. They yield ranged from 25 to 37%, and purity was demonstrated by radioimmunodiffusion, crossed radioimmunoelectrophoresis, polyacrylamide gel electrophoresis, and comparison with other preparations by radioimmunoassay. Immunochemical potency by weight of alphafetoprotein purified from amniotic fluid was similar to that from fetal liver, with molecular weights of 70 000 and 68 500 respectively. The amino acid and carbohydrate composition is also reported. This physicochemical method is relatively simple and inexpensive and is well suited for large scale production.


The effect of the protein content of diluents on peak height in rocket immunoelectrophoresis. We have investigated the effect of different diluents in the quantitation of proteins by rocket immunoelectrophoresis. alpha2-Macroglobulin produced taller peaks as the protein content of the diluent fell to zero, and the dilution was increased. A maximum increase in height of 25-30% was recorded when saline was used as the diluent compared to a sample diluted in sheep serum. Albumin and transferrin showed similar, but less significant effects. Samples and standards should therefore contain approximately equal concentrations of protein for maximum accuracy.


Thyroglobulin of varying molecular sizes with different disappearance rates in plasma following subtotal thyroidectomy. To investigate the possible presence of thyroglobulin (Tg of different molecular sizes in plasma, blood specimens were drawn from patients during and after surgery for thyroid adenoma. Tg was measured in all serum samples by a radioimmunoassay. Selected samples were fractionated on a sepharose CL-6B column, and the fractions were assayed for Tg antigen. In serum drawn at maximum Tg concentration, molecular weights of Tg antigen ranging from 660,000 (19S) to less than 100,000 were found. 6 h later the Tg antigen of mol. wt. less than 100,000 could not be detected, and after 3 days only 19S Tg was present. Correspondingly the plasma Tg concentration vs. time curve showed a biphasic course from which two half-lives could be estimated. For 19S Tg, ta1/2 had a mean value of 4.3 days, whereas the over-all half-life for the mixture of smaller molecules, tb1/2, had a mean value of 3.7 h. The smaller molecules also showed different antigenic reactivity in the assay indicating an altered structure of the molecules. The assumption that this might be due to lack of sialic acid would explain the faster catabolic rate.


The mechanism of basophil histamine release in patients with periodontal disease. Histamine release from washed peripheral blood basophils of thirty-three subjects with varying degrees of periodontal disease was studied. Dental plaque, serum and basophil leucocytes were collected from individual patients. There was no histamine release when autologous, washed sonicated plaque was added to leucocytes. However, the incubation of autologous plaque with serum at 37 degrees C for 30 min generated a factor which induced histamine release from basophils. This serum factor was stable to heat (56 degrees C, 30 min), eluted from a Sephadex G-100 column at a volume corresponding to a molecular weight of approximately 16,000 daltons and its action was inhibited by antibody to C5. This factor, therefore, is probably C5a. There was a variation in the degree of histamine release seen with the leucocytes of different donors. This variability was a property of the basophil rather than a function of the serum. Basophils from patients with gingival indices of 0.5 to 1.0 had significantly more histamine release than basophils from patients with gingival indices of less than 0.5 or greater than 1.5 (P less than 0.001). These experiments demonstrate that dental plaque activates serum to form C5a which in turn releases histamine from basophils. However, these experiments do not indicate a role for IgE in this reaction since the direct interaction of plaque with basophils did not cause histamine release. The release of mediators from mast cells could play an important role in the induction of the inflammatory response in periodontal disease.


The learning of radiological anatomy by medical students. Teaching radiographic anatomy to pre-clinical medical students is important as it relates anatomical studies to clinical medicine and the same time prepares them for the radiology they will encounter in their clinical years. A programme is described in which radiographic anatomy is comprehensively covered without proving too great a commitment to clinical radiologists. The programmes consist of exhibits of X-rays in the dissecting room, slide-lecture demonstrations and a permanent museum of radiological anatomy. This programme has worked well and there has been a very favourable student response.


Lead and other elements in tissues of guinea pigs fed crown vetch grown adjacent to a highway. Forty-four elements were determined in crown vetch sampled along a heavily travelled highway in Virginia. Twenty-four elements including Ba, Br, Cr, Pb, Sn, and Sr were higher in this vegetation than in control vetch. When the highway-grown vetch was fed as 45% (dry weight) of the diet to guinea pigs for 100 days, Ba, Br, Cr, Pb, Sn and Sr were found at higher concentrations in the various animal tissues than in those of guinea pigs similarly fed the control crown vetch.


"Feed a cold, starve a fever"--folk models of infection in an English suburban community, and their relation to medical treatment. This paper outlines a widely-held conception of illness, related to perceived changes in body temperature--'Chills' and 'Colds' on one hand, 'Fevers' on the other---in an English suburban community on the outskirts of London. The relationship between this folk model, and that of the local family physicians is analysed, to show how biomedical treatment and concepts, particularly the germ theory of disease, far from challenging the folk model, actually reinforce it. Remedies which cannot be scientifically and biomedically justified are nevertheless prescribed by the physicians to meet their patients' need to 'make sense' of biomedical treatment in terms of their folk model of illness. At the interface between physicians and patient, biomedical diagnoses and treatment are more 'negotiable' than previously realised--and this has important implications for the delivery of health care.


The use of base pair specific DNA binding agents as affinity labels for the study of mammalian chromosomes. The fluorochromes Hoechst 33258 and olivomycin are base pair specific DNA binding agents. The fluorescence enhancement of Hoechst 33258 and olivomycin in the presence of DNA can be directly related to the A--T and G--C content of the interacting DNA respectively. Cytological observations of metaphase chromosomes treated with these two compounds suggest that the fluorescent banding patterns produced are the reverse of one another.--Non-fluorescent base pair specific DNA binding agents have been used as counterstains in chromosome preparations to enhance the contrast of the banding patterns produced by the base specific fluorochromes. The non-fluorescent G--C specific antibiotic actinomycin-D enhanced the resolution of fluorescent bands produced by the A--T specific fluorochrome Hoechst 33258. Similarly the non-fluorescent A--T specific antibiotic netropsin was found to enhance resolution of the bands produced by the G--C -specific fluorochrome olivomycin. Netropsin was also found to increase the differential fluorescent enhancement of complexes of olivomycin with DNAs of various base composition in solution. These findings suggest that counterstaining agents act through a base sequence dependent inhibition of subsequent binding by base pair specific fluorochromes.--The base specific DNA binding agents have been used to differentiate different types of constitutive heterochromatin in mammalian species, and to facilitate chromosome identification in somatic cell hybrids.


Comparisons of in vivo BrdU labeling methods and spontaneous sister chromatid exchange frequencies in regenerating murine liver and bone marrow cells. BrdU and BrdC have been employed as DNA labeling agents for differentiation of sister chromatids and for extension of sister chromatid exchange (SCE) methods to regenerating murine liver cells in vivo. Comparisons were made between bone marrow and liver cells isolated simultaneously from mice following DNA labeling with either BrdC or BrdU. Although the total mitotic yield of bone marrow cells was considerably greater than in liver, a higher percentage of second division metaphases was observed in liver cell preparations. The percentages of second division c-metaphase cells observed were 31.5% in bone marrow and 73% in liver cell preparations. Utilizing either BrdU or BrdC, no significant difference in percentage of second division metaphases was discerned. The number of spontaneous SCEs per cell was distributed according to the Poisson probability function. No significant differences in mean numbers of SCEs per cell were found in comparisons of bone marrow (1.40) and liver cells (1.65) or of cells which had incorporated BrdU or BrdC.


[Endoscopic findings in Whipple's disease (author's transl)]. In a patient with massive ascites, bouts of fever of unknown aetiology and cachexia, duodenoscopy and laparoscopy revealed white-yellow, partly confluent, nodules and plaques on the duodenal mucosa, and the surface of the liver and the peritoneum of the upper right abdominal quadrant, leading to the diagnosis of Whipple's disease. Histological examinations of these deposits revealed an accumulation of PAS-positive macrophages (SPC cells = sickle-form particle containing cells) and lysosomal bacterial inclusions under the electron-microscope. This raises the possibility of a simple endoscopic method for diagnosing Whipple's disease.


Cortical responses from adults and infants to complex visual stimuli. Event-related potentials (ERPs) time-locked to the onset of visual stimuli were extracted from the EEG of normal adult (N = 16) and infant (N = 23) subjects. Subjects were not required to make any response. Stimuli delivered to the adults were 150 msec exposures of 2 sets of colored slides projected in 4 blocks, 2 in focus and 2 out of focus. Infants received 2-sec exposures of slides showing people, colored drawings or scenes from Disneyland, as well as 2-sec illuminations of the experimenter as she played a game or of a TV screen the baby was watching. The adult ERPs showed 6 waves (N1 through P4) in the 140--600-msec range; this included a positive wave at around 350 msec that was large when the stimuli were focused and smaller when they were not. The waves in the 150--200-msec range, by contrast, steadily dropped in amplitude as the experiment progressed. The infant ERPs differed greatly from the adult ones in morphology, usually showing a positive (latency about 200 msec)--negative(5--600msec)--positive(1000msec) sequence. This ERP appeared in all the stimulus conditions; its presence or absence, furthermore, was correlated with whether or not the baby seemed interested in the stimuli. Four infants failed to produce these ERPs; an independent measure of attention to the stimuli, heart rate deceleration, was demonstrated in two of them. An electrode placed beneath the eye to monitor eye movements yielded ERPs closely resembling those derived from the scalp in most subjects; reasons are given for assigning this response to activity in the brain, probably at the frontal pole. This study appears to be one of the first to search for cognitive 'late waves' in a no-task situation. The results suggest that further work with such task-free paradigms may yield additional useful techniques for studying the ERP.


EEG asymmetry in educationally handicapped children. Bilaterally homologous parietal and temporal EEGs were recorded from two groups of educationally handicapped children while the subjects (Ss) rested with eyes open or closed or performed in simple tasks designed to differentially activate the two hemispheres (reading by Ss, reading to Ss, draw a picture). A 'dyslexic' but not dysphasic group showed a reversal of theta asymmetry from eyes closed to eyes open (L less than R to L greater than R). This difference between groups was reflected most at the parietal placement. Theta in the parietal lead changed in accordance with expectations from reading to drawing (reduction of the R/L ratio), but that pattern did not occur at the temporal lead in 'dyslexics'. A discriminant analysis on theta power correctly classified 20 of the 22 children.


Evoked K-complexes and cardiovascular responses to spindle-synchronous and spindle-asynchronous stimulus clicks during NREM sleep. The hypothesis that the functional role of the sleep spindle is to preserve sleep by inhibiting sensory input (Yamadori 1971) was examined. Series of 44 dB, 10 msec, 1000 c/sec 'clicks' were presented to 12 subjects at a 30-sec ISI during stage 2 sleep either during spindle bursts (i.e. spindle-synchronous clicks) or during interburst periods (i.e. spindle-asynchronous clicks). Contrary to the spindle inhibitory hypothesis, cortical EEG and cardiovascular responses showed no evidence of spindle 'suppression'. Evoked K-complexes were potentiated by the spindle-synchronous stimulation. A second study with 7 subjects replicated this result and extended the finding to include stage 3--4 sleep. It was suggested that the potentiation of evoked K-complexes was due to phasic reductions in inhibitory action during sleep spindles resulting in increased transmission of sensory events or, perhaps, an increase in the lability of certain EEG response systems.


Origin of the neocortically monitored theta rhythm in the curarized rat. An array of epidural electrodes was acutely implanted in locally anesthetized, curarized rats in order to map the surface distribution of rhythmic slow activity (RSA) which appears within the neocortex. Peak amplitudes (of about 122 muV) were centered over the dorsal hippocampus outline. A laminar profile of RSA recorded within the neocortex indicated no shifts in phase relative to a homotopic, epidural electrode. RSA increased slightly in amplitude (mean increase = 53%) at the deepest level or neocortex, but it did not approximate an amplitude peak or null within the neocortex. Multiple-unit activities within the neocortex were not phase-locked to RSA. On the other hand, all of these manifestations of an RSA generator were observed as electrodes passed into the dorsal hippocampus. A unilateral cortical spreading depression (CSD) treatment, which markedly attenuated barbiturate spindles in all subjects (N = 10), usually (N = 7 of 10) had no effect on the neocortically monitored RSA. Dissociations between depressed and non-depressed hemispheres, and between neocortical and hippocampal RSA, were obtained in some subjects during CSD. However, concurrent dissociations were also apparent between recording sites within the hippocampus. It is concluded that the neocortical RSA of rats is passively spread from the underlying hippocampus, and dissociations in neocortical and hippocampal RSA in the rat are secondary to changes in the organization of multiple generators of hippocampal RSA.


Neurophysiological correlates of cognitive development: changes in long-latency event-related potentials from childhood to adulthood. Event-related brain potentials (ERPs) to tachistoscopically flashed slides were recorded from subjects ranging from 6 to 36 years of age. Subjects counted the number of presentations of target slides (P = 0.12) randomly interposed in sequences of background slides (either P = 0.76 or P = 0.88). Also interposed (P = 0.12) in these sequences of targets and backgrounds were one of two types of slides: those (termed dims) bearing any one of the letters from C to Z or those bearing 'novel' patterns, each consisting of a different, quasi-random, 'unrecognizable' color pattern. Targets and backgrounds elicited N1, P2, N2 and P3 waves in subjects of every age tested, suggesting that the mode of processing such explicitly important events and the neuronal substrate for such processing, is similar in children and adults. However, P3 latencies and reaction times in response to these events decreased with age, suggesting that the speed of such processing increases with age. On the other hand, dims and novels elicited ERPs in young children and adults characterized by different late waves. ERPs in 6--8 year olds were characterized by Nc waves (ca. 410 msec and 30 muV) and Pc waves (ca. 900 msec and 30 muV), while ERPs in adults were characterized by P3 waves (ca. 420 msec and 15 muV. The transition from the childhood to adulthood wave form occured in the mid-teens. It is suggested that these differences in ERP wave forms reflect differences in the way children and adults categorize events.


Ethanol dependence in the rat: role of non-specific and limbic regions in the withdrawal reaction. Chronic bipolar electrodes were implanted in cortical, limbic, diencephalic and mesencephalic regions of the rat. Following recovery from surgery the rats were maintained for 14--26 days on a liquid diet in which 35--42% of total calories were provided by ethanol. Following ethanol withdrawal, electrographic and behavioral monitoring was continued for 8--10 h. The withdrawal of ethanol resulted in the time-dependent appearance of a variety of withdrawal signs including tail arching, ataxia, rigidity, tremor and spontaneous and audiogenic convulsions. These behavioral signs were accompanied by the development of epileptiform abnormalities across wide-spread brain regions. Analysis of preconvulsive spike activity revealed a greater spike frequency in limbic, mesencephalic and non-specific diencephalic regions, as compared to those in cortex and specific diencephalon. Seizure discharge during the tonic-clonic phase of the primary audiogenic convulsion was initiated in the mesencephalon or amygdala, but spread rather extensively to the remainder of the brain. In those instances, however, where multiple convulsions occurred following the audiogenic convulsions, there was a marked decline in spread of seizure discharge to the cortex. These results were interpreted to support the notion that some degree of neuroanatomical specificity exists in the genesis of epileptiform abnormalities during ethanol withdrawal. A comparison of these results with those studying the neural mechanisms underlying other forms of generalized epilepsy was made. It is hypothesized that central pacemaking regions such as medial thalamus or reticular formation may serve to organize isolated epileptiform activity into coherent patterns of paroxysmal activity throughout the brain during the ethanol withdrawal syndrome.


Effects of luminance on the pattern visual evoked potential in multiple sclerosis. Pattern visual evoked potentials (PVEPs) were recorded at 5 levels of luminance from 26 patients with multiple sclerosis and from age-matched normal subjects. In normal subjects, the latency of both the major positive peak (P100) and the early positive peak (P60) was an inverse logarithmic function of pattern luminance. The increase in latency per unit log decrease in luminance was 12.1 msec for P100 and 5.7 msec for P60. Only 2 patients had entirely normal results. In 9 patients, the increase in latency of P100 per unit log decrease in luminance was abnormal. Of the 18 luminance-latency functions obtained from testing both eyes, 10 were abnormal, 6 showing a greater than normal increase in latency  with decreasing luminance and 4 a less than normal increase. The 6 luminance-latency functions with a greater than normal increase in latency with decreasing luminance were all from patients without other evidence of optic nerve involvement. Pattern luminance, therefore, as well as patient selection, can significantly affect the proportion of abnormal PVEP latencies in any group of patients with possible, probable or definite multiple sclerosis. The abnormal response of the PVEP to changes in luminance and the different effects upon P100 and P60 indicate that the delayed PVEPs in patients with multiple sclerosis cannot be attributed solely to slowing of conduction in the optic nerve.


Early somatosensory evoked potentials.

Early somatosensory evoked potentials. The early somatosensory evoked potential secondary to median nerve stimulation in the human had an onset latency of 9--12 msec when recorded from scalp electrodes at vertex-to-mastoid, vertex-to-inion or at the base of the skull. Similar latencies were observed from responses recorded over the cervical dorsal columns during neurologic surgery. A latency difference of 1.5 msec was observed between the early response and the responses recorded from the junction of medial lemniscus and nucleus ventralis posterior lateralis of the thalamus during human stereotaxic surgery. Cervical cord transections and transection at the midpontine levels of the monkey showed that the evoked potential was due to generators between these levels. Depth recording of the monkey indicate that the early evoked potential originates in the region of dorsal column nuclei, while the later components are secondary to generators in cerebral cortex.


Auditory brain-stem responses in comatose patients: relationship with brain-stem reflexes and levels of coma. Auditory brain-stem responses (BSR) were recorded in 20 comatose patients in whom the level of brain-stem dysfunction was defined by clinical assessment of brain-stem reflexes and posture. No BSR abnormalities were found in the 10 cases with cortico-subcortical or diencephalic levels. The other 10 patients showed a clear relationship between alteration of the different components of the BSR and the clinical levels of brain-stem dysfunction caused by the rostro-caudal evolution. Alteration of wave P5 seems related to a midbrain dysfunction, of P3 to a pontine dysfunction and of P1 or P2 to a lower brain-stem dysfunction.


Interictal--ictal transition in the feline penicillin epileptogenic focus. A study was undertaken to examine those EEG features which characterize interictal--ictal (II--I) transitions in the feline penicillin focus. The study describes certain complex changes in spike frequency and their statistical analysis, which appear to have predictive value in signaling II--I transitions. The significance of these data with respect to the conflicting results of other studies and their possible application to clinical epilepsy are discussed.


A flexible low cost signal averager.

A flexible low cost signal averager. A 2 channel signal averager based on the KIM-1 microcomputer module is described which can be easily assembled for under $500. Number of samples per pass can be set to 128 or 256 and sample time is variable from 1 msec to 256 msec. Averaged data can be displayed on an oscilloscope, output slowly for use of a chart recorder, or stored on a low cost audio cassette recorder for later analysis. Simply adding additional memory (for less than $100) allows it to be expanded to 8 channels.


Digital memory recorder in electromyography and nerve conduction studies. Specifications are given: (i) for a 3-channel digital memory that, when connected to an inkjet writer, allows action potentials from muscle and nerve to be recorded without distortion (upper limiting frequency 10,000 Hz); (ii) for a 14-channel digital memory connected to a 14-channel inkjet writer for the measurement of the territory of the motor units by recording simultaneously from 14 leads of a multi-electrode spaced over the cross-section of the muscle; (iii) for a digital circuit that rejects signals that exceed a given amplitude and duration and interfere with electronic averaging of potentials from sensory nerve less than 1 muV in amplitude.


A clinically effective spike recognition program: its use at electrocorticography. This automated system detects spikes when the first derivative value of the EEG or ECoG signal exceeds a user-determined threshold twice, with reversal of direction, within 30--80 msec. The EEGer selects this threshold by his assessment of the system's initial performance in a recording situation. By combining the spike detecting abilities of both the EEGer and computer, two tasks can be accomplished on-line for 16-channel EEG or ECoG: (1) localization of the origin of widely synchronous spikes, and (2) assessment of the relative quantities of spikes emanating from several active regions. This system achieved a human-computer correlation ranging from 0.79 to 0.84.


Bereitschaftspontential during the acquisition of a skilled motor task. Bereitschaftspontential (BP) research typically uses very simple, abrupt movements that require no learning and which change little over the duration of the experimental session. The present study investigated changes in the size and cortical distribution of the BP during the acquisition of a skilled motor task. Twelve subjects were employed. Electrodes at FZ, CZ, C3'' and C4'' were used to record the EEG with DC amplifiers. A series of 6 button presses in a specified pattern constituted the motor task. Subjects were instructed to press the series every 20 sec as quickly as possible, but with no errors. Significant response time, electrode and trial main effects, and electrode by trial interactions were found. The BP increased steadily at all electrodes as performance improved, i.e., as response time decreased. After the response reached asymptote the BP recorded at FZ and C4'' decreased, while the BP at CZ and C3'' remained relatively constant. Multivariate analyses of covariance showed a consistent relationship between the improvement in response times and the changes in the BP. This study demonstrates that the size and cortical distribution of the BP are systematically related to improved proficiency of a motor response with learning.


Auditory brain stem potentials with alcohol. Auditory brain stem potentials were recorded from unrestrained rats and from cats paralyzed with Flaxedil, before and after ingestion of intoxicating dosages of alcohol. The acute effect of alcohol was a cumulative increase in the central conduction times of successive brain stem potential peaks. Statistically significant latency changes were found for peaks attributed to neural structures as far peripherally as the medulla. This depressive influence of alcohol on sensory transmission was independent of variations in stimulus intensity and body temperature.


Activity of human hippocampal formation and amygdala neurons during memory testing. Single and multiple unit recordings were made from fine wires stereotaxically implanted in the hippocampus (HC), hippocampal gyrus (HCG), and amygdala (Am) of psychomotor epileptics. During a series of memory and control tests presented on slides, 21 of 155 HCG units, 15 of 59 HC units, and 2 of 54 Am units showed what appeared to be simple phasic or tonic visual responses. Twenty-seven other units, found only in the HCG, changed firing only during slides requiring a choice ('choice units'). A given choice unit responded during choices indicated verbally or manually, and during tasks requiring recall of Recent Memory, various visual discriminations, and expressions of preference. Choice units were not affected by sensory stimulation or motor activity in contexts not requiring choice. Phasically inhibited choice units had higher firing rates and lower signal-to-noise ratios than tonically excited units. Whether an electrode recorded a choice unit was unrelated to if it recorded a response to hyperventilation, or was in an area of epileptic pathology. Recordings were also made during an interview lasting several hours and eliciting a wide range of behaviors. Five of the 131 HCG units fired in repeated extended bursts, at least 50 times background during recall of word pairs or of the patient's hospital room. The unit response did not occur during numerous control tasks possessing similar overt sensory, motor, and social concomitants, but not requiring Recent Memory.


Experimental kuru in the rhesus monkey: a study of EEG modifications in the waking state and during sleep. EEG patterns recorded in the waking state and during sleep were studied in 6 rhesus monkeys inoculated with a strain of Kuru previously passaged in rhesus monkey (ENAGE strain, rhesus L6 56). The onset of the disease was confirmed by the appearance of various clinical signs in 4 monkeys 15 months after inoculation. At the 16th month, the first EEG modifications appeared during sleep, which became lighter. The waking EEG was abnormal during the mature phase of the disease; it was characterized by slow anomalies and scattered spikes. The sleep EEG still presented 3 stages of Slow Wave Sleep which, however, were totally unlike the physiological stages. REM sleep rapidly disappeared, as did the cyclic organization pattern. Irritative phenomena became very significant and, in particular, very frequent 'tonic seizures' were observed. Experimental Kuru thus appears, in the rhesus monkey, as an epileptogenic encephalopathy, which is differentiated from both the human disease and the experimental disease in the chimpanzee.


Excessive daytime sleepiness in man: multiple sleep latency measurement in narcoleptic and control subjects. Excessive daytime sleepiness is a complaint characterizing many disorders of the wakefulness--sleep cycle. This paper addresses the complaint of sleepiness objectively by an attempt to differentiate a group of control subjects from a group of patients with unambiguous narcolepsy. Fourteen control and 27 narcoleptic subjects were evaluated by one of three protocols involving nocturnal recordings, detailed interviews, and 5 or more 20-min opportunities to sleep offered at 2-h intervals beginning at 10.00 o'clock, +/- 30 min. Each 20-min opportunity to sleep was given to subjects lying in a darkened quiet room and asked to try to fall asleep. Polysomnographic variables were monitored and sleep was scored in 30-sec epochs by standard criteria. The interval from the start of each test to the first epoch of NREM (including stage 1 sleep) or REM sleep was called sleep latency. In two of the protocols, the subjects were awakened immediately after sleep onset. In the third protocol, the subjects were awakened after 10 min of sleep. Narcoleptics consistently fell asleep much more readily than did control subjects. We conclude that the Multiple Sleep latency test, in addition to providing opportunities to clinically document sleep onset REM sleep periods, can demonstrate pathological sleepiness. Based on these data, we suggest that an average sleep latency less than 5 min be set as the minimum cutoff point for pathological sleepiness.


Size-specific information channels and selective attention: visual evoked potential and behavioral measures. Subjects were shown a stimulus (diffuse light or a checkerboard pattern with checks subtending 12' or 35' of arc) and were instructed to remember and attend (give a reaction time response to or count) that stimulus during the course of an approximately 6-min trial. A trial consisted of the random presentation of 8 stimulus flashes (diffuse light and checkerboards with 9, 12, 18, 24, 35, 45, 94 and 95 min checks) at a rate of 1/555--930 msec. Visual evoked potentials and reaction times were averaged to each of the 8 stimuli. The purpose of the experiment was to assess how specifically the relevant stimulus could be attended, as indicated by VEPs and behavioral responses to the relevant and 7 irrelevant stimuli. Attention to one check size resulted in greater amplitude VEP components to that size than to other sizes: the greater the discrepancy between the flashed and attended check size, the smaller the VEP amplitude. Such size tuning was first evident in the VEP 160 msec after stimulation and most evident at 260 msec. The bandwidth of the VEP amplitude size-tuning functions became progressively narrower from 160 up to 260 msec, that point in time when the RT response was initiated. The width at 260 msec was similar to that indicated by the behavioral RT data (modal latency of 322 msec). The functional components of the VEP appeared to reflect the effects of attention on the activity of cortical size channels or detectors.


Interhemispheric synchrony in the EEGs of full-term newborns. Cross correlations between 4 homologous pairs of ear-reference derivations were calculated for 45-sec digitized samples of typical active and quiet sleep in 19 selected normal newborns at 21--70 h after term birth. Repeat recordings were obtained in 9 subjects 24 h after the original recordings and another set of cross correlations was calculated. For comparison, similar recordings were obtained from 4 asymptomatic young adults and cross correlations were calculated for 45-sec samples of typical stage 4 and stage REM sleep and for stage W both before and after sleep. Mean cross-correlation coefficients at zero time lag were generally higher for the adult sleep data than for the newborn data, but the differences were smaller than had been anticipated. The coefficients for active sleep data in the newborns were higher than for quiet sleep data, while in the adults the coefficients were higher for stage 4 than for stage REM. Coefficients were higher for convexity derivations than for temporal derivations in both newborns and adults. A review of the small available literature suggests that recording and analysis techniques may be significant variables determining the results obtained in such studies. The cross-correlation method of measuring interhemispheric synchrony may prove useful in identifying brain disorders in the postnatal period.


A study on the Valsalva manoeuvre in young healthy subjects. The Valsalva manoeuvre was performed by 2 groups of young healthy male subjects: novice divers and professional divers. The electrophysiological features of this manoeuvre are summarized and the results of the test are correlated with subject age, motivation and FEV1/VC coefficient (forced expiratory volume sec/vital capacity). Positive test results (fainting caused by the Valsalva manoeuvre) increased with subject motivation (novice divers), FEV1/VC coefficient and decreasing subject age. The various hypotheses proposed to account for fainting caused by the Valsalva manoeuvre are discussed with respect to these data. The implications of Valsalva positivity in young divers are discussed.


The effects of local micro injections of opiates and enkephalins into the forebrain on the electrocorticogram of the rat. The effects of various opiate compounds have been studied on the electrocorticogram (ECoG) of the rat following local injection into various brain areas. Injections of all compounds studied into both the caudate-putamen and the basal forebrain, in particular the olfactory tubercles, induced changes in the ECoG. Injections of saline vehicle into these areas were ineffective as were injections of morphine into the corpus callosum. Potency was etorphine greater than morphine = codeine greater than thebaine. Naloxone alone was inactive following injection but if combined with morphine markedly attenuated the normal morphine response and reversed the morphine response if injected following morphine. The endogenous opiate compound enkephalin and the synthetic analogue D-ala2-met5-encamide also induced electrocortical changes which were naloxone sensitive. The results are similar to those following systemic administration of opiates. It is possible that the areas studied represent the site of action of systemically applied opiates. It is suggested that the opiates and enkephalins produce their actions by acting at the same site. Since the areas studied are rich in dopaminergic terminals an interaction may exist between dopaminergic and opiate mechanism to bring about the observed changes.


[Electrophysiological study of latent neuropathies induced by perhexiline]. The aim of this study was to estimate the frequency of latent perhexiline-induced neuropathy in man. Several electrophysiological parameters were recorded and compared between a control group and a group of treated patients, without clinical evidence of neuropathy. The Hmax/Mmax amplitude ratio of the soleus H reflex was decreased in 2/3 of the patients. The H reflex latency was increased in 1/3 of them. The distal motor latencies of ulnar and popliteal nerves were increased in 1/4 of them. Only 1/3 of the patients treated did not show any electrophysiological abnormality. The validity of these results and the possible mechanisms involved are discussed.


Auditory evoked potentials and transcendental meditation. Auditory evoked potentials to tone stimuli were recorded from 8 practised meditators before, during, and after meditation, and also during light sleep. No consistent changes were noted between baseline and meditating AEPs, or between meditating and sleep AEPs.


Long term antithyroid drug therapy for intractable cases of Graves' disease. Among the 504 patients with Graves' disease in whom the author initiated antithyroid drug therapy between 1956 and 1968 are 14 to whom the administration of antithyroid drug has been continued for 8 to 21 years because of recurrence shortly after the discontinuation of drugs. During these periods, no side effects were observed and three patients went into complete remission after 8, 9 and 20 years of antithyroid drug administration, respectively. It is suggested therefore that the long term administration of antithryoid drugs can be a useful treatment for intractable cases.


Reaction of tRNAPhe from yeast with 1-fluoro-2,4-dinitrobenzene. Attachment sites of the potential antigenic-determining 2,4-dinitrophenyl residues. The reaction of 1-fluoro-2,4-dinitrobenzene with tRNAPhe from yeast, for the introduction of antigenic-determining 2,4-dinitrophenyl residues into tRNA, took place only at adenosine residues in tRNAPhe. After reaction at pH 8.0 and 50 degrees C two kinds of products were detected: one was ribose-modified adenosine which was derived from the 3' terminus of tRNA, and the other was base-modified adenosine. The sites and extent of the modification of each particular adenosine residue of tRNAPhe were determined as follows: 5 (6% modified), 31 (2%), 35 (36%), 67 (5%), and 76 (51%). Thus mainly the terminal adenosine and one adenosine in the anticodon loop bear the 2,4-dinitrophenyl residue.


Comparison of the RNA-dependent DNA polymerase of an endogenous avian leukosis virus to the polymerase of an exogenous avian leukosis virus. RNA-dependent DNA polymerases from Rous-associated virus-O and avian myeloblastosis virus were partially purified by affinity chromatography and compared to each other. The enzymes are indistinguishable in the immunoglobulin inhibition test and by several enzymological criteria, such as optimum curves for the concentrations of Mg2+, K+, H+; monophasic Lineweaver-Burk plot for dTTP and biphasic Lineweaver-Burk plot for dGTP. In thermal inactivation studies a small difference can be observed, suggesting a minor difference in the physical structures of the enzymes. Our findings are consistent with the idea that the RNA-dpendent DNA polymerases of endogenous and exogenous avian leukosis viruses are very closely related to each other and therefore may be regarded as one group of polymerases.


The DNA . tyrocidine complex and its dissociation in the presence of gramicidin D. The peptide antibiotic tyrocidine affects transcription in vitro by interfering with the initiation process. The complex formed between tyrocidine and native DNA is stable against nucleolytic enzymes. It is sensitive to variations in the salt concentration. Protection of DNA as a function of the tyrocidine concentration follows a curve suggesting cooperative binding of tyrocidine to DNA. The complex formation at low tyrocidine concentrations is accompanied by the presence of single stranded regions of unknown length. With single-stranded DNA, tyrocidine forms a complex which is relatively insensitive to high salt concentrations. Both native and single-stranded DNAs of the complexes become digestible in the presence of gramicidin D, another peptide antibiotic. Breakdown of the complex as a function of the gramicidin D concentration follows a curve which is the reverse of cooperative binding. Gramicidin D acts more effectively on single-stranded rather than on double-stranded DNA . tyrocidine complexes. The DNA . tyrocidine complex, which is broken down by formamide and dimethyl sulfoxide is stabilized by gramicidin D. The implications of these results are discussed with respect to a possible recognition of DNA sequences by tyrocidine and the interaction between DNA, tyrocidine and gramicidin D.


CSF circulation and blood-CSF barrier.

CSF circulation and blood-CSF barrier. Results of microzone electrophoresis of non-concentrated CSF after staining with nigrosine and after evaluation on non-transparent acetate film are compared with those of isotope cisternography (111In-DTPA). We found that blood-CSF barrier disturbances begin with an increase of the absolute values of prealbumins in normal CSF circulation. When a barrier impairment occurs, by first increasing the absolute values of alpha1-globulins, we state a pathological CSF circulation. In this case, most of the globulin region is pathologic (globulin-type).


Air cystometrography and sphincter electromyography in patients with bladder outlet obstruction. 38 patients with infravesical obstruction secondary to benign prostatic hypertrophy had CO2 cystometrography (CMG) and sphincter electromyography (EMG) at onset of retension and 6 weeks after intermittent bladder drainage. Neurological diseases were excluded in these patients. Three patterns of CMG tracings were obtained and were called, normal, 'compensating' and 'decompensating' on the basis of the volume: pressure ratio of the curves. By strict definition, no abnormal detrusor contractions were seen in any of these patients who also all had normal sphincter electromyography. It is questionable therefore if infravesical obstruction, in the absence of neurological deficits can account for the uninhibited detrusor contractions described in the literature. Rather it is possible to explain the above CMG tracings on the effects of obstruction on the bladder smooth muscle and collagen fibers which have been shown to be non-neural dependent.


Immune response against the beta-galactosidase enzyme of E. coli at precursor cell level. I. Analysis of the secondary repertoire in BALB/c mice. The BALB/c secondary response against the beta-galactosidase (beta-gal) enzyme of E. Coli was analyzed at the precursor cell level by using the splenic focus technique. Our results indicate that in immunized mice, one out of 18 000 B cells is able to recognize beta-gal. Among the families of anti-beta-gal monoclonal antibodies, a subset of specific antibodies was detected which is capable of protecting the enzyme from heat denaturation. The frequency of clones making protecting antibodies is 1 out of 90 000 and appears to be fairly constant among different individual mice. Further, the degree of heterogeneity of protecting antibodies analyzed in one individual is very high (250-fold difference in affinity) but comparable to other secondary repertoires. Specific frequencies are compared with previous findings relative to secondary responses against artificial haptens. It is suggested that a different type of recognition exists between protein determinants and artificial haptens. In addition, the relatively high proportion of clones making antibodies of the protecting type suggests that only a small proportion of antigenic sites on the beta-gal is actually able to stimulate an immune response.


Detection of delayed hypersensitivity reaction in rats by radioisotopic footpad assay with sodium deoxycholate extract and mitomycin-C-treated tumor cells. Immune response in WKA rats immunized with methylcholanthrene-induced KMT-17 tumor was measured by radioisotopic footpad assay. The assay was specific, quantitative, and objective compared with the ordinary method of measuring the thickness of footpads. The strongest footpad reaction was observed 24 hr after injection of the antigens. The reaction was transferred by lymphoid cells but not by serum. These results indicate that the footpad assay manifests delayed-type hypersensitivity to tumor antigens. In order to obtain more reliable antigen preparation for the footpad assay, antigens prepared by several methods were compared. Solubilized antigen extracted with sodium deoxycholate (DOC) showed the strongest reaction in the immunized host and weak reaction in the control non-immunized host. This marked difference between the immunized and control groups indicates that DOC-extracted antigen was better antigen than tumor cells treated with mitomycin-C (40 microgram/ml), formaldehyde solution (0.2%, 0.01%), X-irradiation (3,500, 10,000 rad), non-treated whole-cell antigen, crude membrane, cell homogenates, or antigens extracted by hypotonic sonication and 3M KCl method. The DOC-extracted antigen was well preserved at -20 degrees for 1 month.


Effect of protease inhibitors in mice treated with the hepatocarcinogen, hexachlorocyclohexane (alpha-isomer) and the bladder carcinogen, N-butyl-N-(4-hydroxybutyl)nitrosamine. Studies were made on the effect of protease inhibitors (pepstatin, leupeptin, and antipain) on the induction of tumors in mice by the alpha-isomer of 1,2,3,4,5,6-hexachlorocyclohexane (alpha-BHC) and by N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN). Animals were given 0.05% alpha-BHC in the diet or 0.05% BBN in their drinking water and then fed on a powder diet supplemented with 0.1% protease inhibitors. The incidence of liver nodular hyperplasias was significantly higher in the group treated with alpha-BHC and leupeptin than in the group treated with alpha-BIC alone, but pepstatin and antipain did not affect induction of liver nodular hyperplasias by alpha-BHC. None of the three protease inhibitors influenced the induction of bladder tumors by BBN.


Leukocyte migration inhibition induced by the combination of drug and a liver constituent in patients with drug-induced hepatitis. The leukocyte migration inhibition test in agarose medium was performed in 23 cases of clinically diagnosed drug-induced hepatitis. When the test antigen was the combination of soluble phase of a liver homogenate fractionated by Sephadex G-100 which should have contained liver specific antigen and the offending drug the leukocyte migration was inhibited in 86% of cases. Whereas none of 12 cases of drug allergy without hepatic injury showed a positive result with the same combination of antigens. Other organ homogenate-muscle and kidney-never gave positive results when mixed with the offending drugs in cases of drug-induced hepatitis. It was concluded that in hypersensitivity type drug-induced hepatitis cell-mediated immunity might be established to the complex of liver specific antigen and the drug.


Effect of glucagon infusion on the renal clearance of amylase relative to creatinine. Recent data seem to support a tubular defect as the mechanism of the elevated renal clearance of amylase relative to creatinine in acute pancreatitis. Glucagon has been proposed by some to be an important factor in this phenomenon. To examine the role of glucagon as this "tubular dysfunction factor", we investigated the effect of intravenously infused glucagon on the fractional excretion of amylase and the tubular handling of a low molecular weight protein, beta2 microglobulin, in normal, healthy volunteers. At glucagon levels far in excess of those seen in pancreatitis, the clearance ratio of beta2 microglobulin relative to creatinine increased, whereas the clearance ratio of amylase relative to creatinine did not increase above the normal range. The dissociation between beta2 microglobulin clearance and amylase clearance allows one to question the theory that tubular dysfunction is the mechanism of the elevated renal clearance of amylase relative to creatinine in acute pancreatitis. Glucagon does not appear to be the sole factor responsible for the elevation of renal clearance of amylase relative to creatinine in acute pancreatitis.


Muscular equivalent of the lower esophageal sphincter. In order to measure muscle thickness and to define the muscular architecture at the gastroesophageal function, both en bloc fixation and a new method of preparing dried fiber specimens were used. Specimens were obtained from 32 kidney donors and human cadavers. Wall thickness was measured at 32 identical locations in the esophagus and stomach. The oblique gastroesophageal ring (GER) was the site of greatest muscular thickness and served as a reference point. From the GER the muscle thickness tapered (P less than 0.05 to P less than 0.001) in both a cephalic (esophageal) and caudal (gastric) direction for a length of 31 mm +/- 2.5 SD. The increase in thickness was due to an increase in the muscle mass (fiber aggregation) of the inner muscle coat. The muscle bundles of this coat split up 10.2 mm +/- 3.0 SD above the GER (fixed specimen) and for a length of 25 mm +/- 8 SD formed short transverse muscle clasps on the lesser curve side. Those muscle bundles on the greater curve side formed long oblique gastric fiber loops. The angle of His was inconstant in location and distal to the uppermost gastric oblique fibers (18 mm +/- 7 SD) and to the GER (9 mm +/- 6 SD).


Nuclear binding of tritiated actinomycin in surface epithelial cells from normal stomach and atrophic gastritis. Triated actinomycin binding to DNA is closely linked to the degree of repression in chromatin. 3H-AM binding to DNA is the most pronounced in nuclei of cells committed into cycle. Inversely, in cells in the last steps of their differentiation or (and) in the resting state (non-dividing cells), 3H-AM binding for DNA is diminished down to a baseline since it is limited by the deoxynucleoproteins. Epithelial cells of stomach mucosa and duodenum demonstrate an increased cell uptake of tritiated actinomycin from the surface to the bottom of the pits. In severe gastritis and in intestinalysed metaplasia this was abolished: with a uniform enhancement of 3H-AM binding. These findings seem to indicate that these cells are derepressed.


Immunohistochemical demonstration of several tumour markers in neoplastic and preneoplastic states of the uterine mucosa. Enzymic immunocytochemical demonstration of five tumour markers (CEA, AFP, kappa-casein, total HCG and its beta-subunit) was performed on various pathological states of the uterine mucosa. CEA was constantly found in areas of atypical hyperplasia and in more than 60% of cancers. kappa-Casein on the contrary was an unexpected but frequent finding in cystic hyperplasia. The other antigens were not, or exceptionally, detected in the tissues. We believe that CEA detection may be a very useful tool in delineating preneoplastic changes of the endometrium while the presence of kappa-casein "storing cells" might point to an unopposed estrogen action.


[Percutaneous transhepatic intubation of bile ducts for combined internal and external drainage in extrahepatic cholestasis (author's transl)]. In 37 patients with extrahepatic cholestasis of different origin, percutaneous transhepatic intubation of the bile ducts was performed for combined internal and external drainage. In 5 cases with stones in the extrahepatic bile ducts and in 12 cases with periampullary tumors, the bile ducts were drained preoperatively. In 3 of these patients, temporary drainage was performed because of postoperative complications. Thirteen patients with non-resectable tumors received a permanent drainage as palliation. In 4 patients temporary drainage was performed during radiation/cytostatic therapy. One patient with an iatrogenic bile duct lesion and one patient with an inflammatory tumor in the liver hilum had a drainage catheter placed because a bile duct resection with creation of a biliodigestive anastomosis was not possible. In one patient with a non-resectable periampullary carcinoma the drainage catheter was replaced by a teflon bile duct endoprosthesis inserted by the percutaneous transhepatic approach. The complications of non-operative percutaneous transhepatic bile drainage are discussed.


[Size distribution of colonies for the detection of cytogenetic lesions due to X-rays and cytostatics]. 1. The "all or nothing" principle in the colony survival test has to be given up. By reproductive death of mammalian cells we can conclude that colonies with two and more cells in the colony survival must be considered. An arbitrary drawn low limit of cell numbers can give misleading results. 2. The colony-size spectrometry registers the reproductive lethal cell damages. These cell damages could be detected qualitatively as well as quantitatively. The sensitivity of this method is greater on low and middle X-ray dose as compared with the colony-survival test. 3. Sublethal damages with tendency of recovery as produced by the split-dose effect are not observed by colony-size-spectrometry. Therefore, this damage is not due to chromosomal X-ray-damages. 4. The combination of physical and radiogenic damages as well as the combination of radiogenic and pharmacological damages and lethal reproductive cell damages also can be seen. The colony-size-spectra observed show that only additive but no potentiating effects occur. 5. There is a linear relationship between MKD and X-ray dose. This relationship becomes asymptotic using cytostatics as in the case of Bleomycin and ICRF 159 in a dose range beyond approximately 2 mg/kg.


Isolation and some properties of a new enzyme-binding protein in rat plasma. alpha-1-Inhibitor3 (alpha-I3), a new enzyme-binding protein, was isolated from rat plasma by a combination of ammonium sulfate precipitation, ion exchange chromatography on DEAE cellulose and gel filtration on ultrogel AcA34. Agarose gel electrophoresis of the purified inhibitor showed a single protein band with alpha1-mobility giving a single precipitation line on immunoelectrophoresis against anti-rat serum. A specific antiserum against the purified inhibitor was raised in rabbits. alpha1-I3 showed immunologic cross-reaction with human inter-alpha-trypsin inhibitor. alpha1-I3 formed a complex with trypsin, which was thereby inhibited; the electrophoretic mobility of the complex was less than that of free inhibitor. Inflammation, induced by turpentine, caused a decrease in the serum concentration of alpha1-I3 to 36% of the initial value within 48 h. alpha2 acute phase macroglobulin (alpha2-AP) showed a simultaneous increase to 7.1 g/l and alpha1-antitrypsin (alpha1-AT) to twice its normal value.


The potential value of immunohistologic techniques in the classification of ovarian and testicular tumors. The diagnosis of neoplasia continues to rest upon the judicious application of morphologic criteria for the identification of tumor cells. Immunohistologic techniques offer the pathologist an alternative means of cell identification according to the antigenic constitution of the cell or its products. The potential effect of these techniques upon the diagnosis and classification of neoplasia is illustrated by the application of immunoperoxidase methods to the study of alpha-fetoprotein, human chorionic gonadotrophin, and steroid hormone localization in tumors of the ovary and testis.


Ag staining of the nucleolus organizer (NO) and its relationship to satellite association. The frequency of involvement in satellite association and the frequency of selective staining of the secondary constrictions with silver solutions have been studied in five phenotypically normal individuals, all carriers of morphological variants of the nucleolus organizing region (NOR). The results show the preferential involvement of some morphological markers in satellite association, and also their preferential staining with Ag-I. It has also been shown that acrocentric chromosomes involved in satellite association are always stained by silver.


Differences in binding affinity of human IgE for receptors in chopped human lung. Experiments were performed to ascertain whether IgE in different allergic sera had the same or different sensitizing properties for chopped human lung. When allergic sera were allowed to compete with myeloma IgE for tissue receptors in chopped lung and subsequently challenged with antigen, two groups of sera could be distinguished, one which competed well with myeloma IgE and one which competed poorly. Sera that competed well with myeloma IgE were also able to sensitize for greater histamine release relative to IgE concentration when sensitized lung tissue was challenged with anti-IgE. The converse was true of those sera that competed poorly with myeloma IgE in the antigen assay, in that they sensitized for histamine release only at relatively high IgE concentrations in the anti-IgE assay. The possible significance of these findings is discussed.


Tissue antigens in seminal secretions. I. Precipitin reactions of rabbit antiserum to human seminal plasma. Precipitating antibodies prepared in rabbits with human seminal plasma (HSP) have been used in a preliminary study to define the antigenicity of HSP by use of gel precipitin methods. Also, a method for evaluating and preserving liquid precipitates of acid phosphatase in polyacrylamide is described. Some ten or eleven antigens were demonstrated by Ouchterlony agar immunodiffusion (ID). Eight of these appeared to be HSP-specific and two or three were also found in normal human serum (NHS). Five of the HSP-specific antigens were shown by agar immunoelectrophoresis (IEP) and by ID to be acid phosphatase. Four of these constituted the basic isoenzyme pattern and were shown by IEP and ID to be shared by prostatic fluid (HPF). Titration of HPF by ID yielded antigen titres of 46, 33, 17 and 3 for the four isoenzymes. IEP patterns obtained with HSP and HPF showed two isoenzymes migrating to the alpha-globulin zone. The third isoenzyme migrated to the alpha-2-fast beta area, while a fourth was seen as a slow beta precipitin arc formed consistently by HPF, and variably by HSP. HSP contained a fifth isoenzyme which also migrated to the slow beta area. Human prostate extract (HPE) showed only two, sometimes three, of these components, one of the slower components being frequently faint in appearance or absent. In HSP and HPF isoenzyme migrating to the alpha-2-fast beta zone exhibited two distinct characteristics: biphasic migration and limited diffusion, suggesting a heterogeneous high molecular weight isoenzyme or an isoenzyme complex. However, observed differences between HSP, HPF and HPE in migration rate, relative concentrations of subfractions, lateral diffusion, etc. leave open the question of the nature of the heterogeneity.


Investigations on strain-specific and common antigen antihemagglutinin antibodies oriented to the influenza virus H3N2 strains A/Hong Kong/1/68 and A/Port Chalmers/1/73. Antisera raised by vaccination of chickens with the H3N2 influenza virus strains A/Hong Kong/1/68 and A/Port/Chalmers/1/73 were found to contain common antigen antibody (CAA), combining with both viruses, and strain-specific antibody (SSA), combining with homologous virus only. The reaction of CAA with homologous and heterologous viruses and of SSA with homologous virus was investigated by means of equilibrium filtration. CAA was found to be antibody of higher quality than was SSA. The virus strains tested were found not to contain identical common antigenic determinants, and CAA and SSA competed for the same antigenic sites when allowed to react simultaneously with homologous virus. The kinetics of reaction of CAA and SSA with the corresponding viruses was tested by means of the photometric antibody concentration unit method. It was found that SSA and CAA of a given strain have the same specificity and that the reaction of CAA with heterologous virus is registered as heterologous antigen-antibody interaction. These findings suggest that SSA and CAA are antibodies of the same specificity but with differences in quality.


Antitumor activity of mycobacterial glycolipid A1. Glycolipid A1 isolated from Mycobacterium bovis BCG, when dissolved in olive oil and injected together with Line 10 transplantable hepatoma cells, is able to elicit a host response which results in the abrogation or retardation of tumor growth in syngeneic guinea pigs. Glycolipid A1 does not have adjuvant activity for delayed type hypersensitivity, and antibodies to A1 have not been detected in the sera of guinea pigs during or after the tumor abrogation induced by A1 injection Glycolipid A1 does not share antigenic determinants with Line 10 cell lipid fractions. The possible role of the granuloma response elicited by A1 in controlling tumor growth is discussed.


Immune interferon induced by phytohemagglutinin in nude mouse spleen cells. Phytohemagglutinin is able to trigger interferon synthesis in spleen cell cultures from nude (nu/nu) mice as effectively as in splenic cell cultures from haired, control (nu/+), thymus-bearing mice. A minor theta-bearing cell population present in the spleen of nude mice appears essential to phytohemagglutinin interferon production, although cooperating cells are also required. The properties of nude mouse phytohemagglutinin interferon are indistinguishable from those displayed by the interferon induced in thymus-bearing mouse spleen cell cultures. Both interferons are unstable at pH 2 and cannot be neutralized by an antiviral interferon serum; hence, their characteristics correspond to those described for type T interferon. As in the case of viral interferon, pretreatment of L cells with nude phytohemagglutinin interferon induced specific enhanced phosphorylation of a 67,000-molecular-weight protein in vitro when cell extracts were incubated with double-stranded RNA and gamma-[32P]ATP.


Antigens of pig follicular fluid and ovarian tissue and the effect of anti-ovarian serum on pig oocytes and spermatozoa. Common antigens were detected in pig follicular tissue, blood serum, blood plasma, seminal plasma and kidney, liver, testicular and ovarian tissue. If pig oocytes were exposed to the action of anti-ovarian antibodies, no precipitation of the zona pellucida was observed. In immunofluorescence, however, reaction of the zona, which was unaffected by absorption of this antiserum by any of the above antigens, was found. Antisera against pig spermatozoa, seminal plasma, follicular fluid, and testicular tissue did not react with the antigens on oocytes. Similarly, anti-ovarian and antifollicular sera had no effect on boar spermatozoa.


Dissociation, aggregation of sesame alpha-globulin in nonionic detergent solution. Nonionic detergents Triton X-100 and Brij 36T induce dissociation and aggregation of the protein sesame alpha-globulin above the critical micelle concentrations (cmc) of the detergents. Spectrophotometric titration in Triton shows no change in the pKInt value of the tyrosyl groups at 1x10-3 M detergent where both dissociation and aggregation of the protein are observed. Fluorescence measurement does not indicate any change in the environment of the tryptophan groups of the protein in Brij. Viscosity measurements show no major conformational change of the protein in the detergent solution. Binding measurements suggest that perhaps micelles of the detergent predominantly bind to the protein. The detergent micelles preferentially bind to the exposed hydrophobic surfaces of the protein subunits. The association of the protein detergent complex through electrostatic interaction is probably responsible for the formation of the aggregates.


Motor innervation of the urinary tract studied by retrograde axonal transport of protein. A new method for tracing the nervous control of the bladder is described. A protein substance, horseradish peroxidase, has been shown to be absorbed and transported by nerve fibers. The substance was injected into the bladder neck region of a series of dogs. After uptake by motor neurons the horseradish was carried to the spinal cord segment controlling micturition. Implications regarding the neurophysiology of the urinary tract are discussed.


Establishment and characterization of a new human urinary bladder carcinoma cell line (PS-1). An epithelioid cell line (PS-1) has been established from a transitional cell cancer derived from human urinary bladder. Subcutaneous injection of the epithelioid cells into weanling athymic nude mice induced solid tumors histologically similar to the original tumor. A cell line was also established from a tumor induced in the athymic nude mouse (PS-1, T-1). Both cell lines exhibited essentially identical growth characteristics and formed a monolayer growth of epithelioid cells in culture. Electron microscopic studies confirmed epithelioid morphology. No fibroblastoid elements were observed. Chromosomal analysis revealed heteroploidy with persistent marker chromosomes; all cells contained a Y chromosome. The presence of tumor-specific antigen(s) in PS-1 cells was suggested by microcytotoxicity assays with peripheral allogeneic lymphocytes from other transitional cancer cell patients. Sera of urinary bladder cancer patients reacted with nuclear antigens of the established cells.


Specific inhibition of XC syncytia formation by anti-MuLV gp69/71 antibodies. The XC syncytia formation observed when ecotropic murine leukemia virus producer cells were cocultivated with XC cells was specifically inhibited by anti-MuLV gp69/71 antibodies with type- or group-specific activities. Thus, the major glycoprotein of budding viruses was probably responsible for the fusion with XC cells. Since this inhibition is specific for anti-gp69/71, it provides a simple, sensitive and convenient way to type ecotropic type C viruses and to detect anti-gp69/71. The micromethod we have developed has been applied to the detection of type-specific anti-GLV gp69/71 natural antibodies in the serum of normal mice.


[Sperm allergy of the anaphylactic type]. A case of an anaphylactic type of allergy is reported with pruritus, mucosal edema, asthma and shock symptoms which developed in a 26 year old female against her husbands sperma fluid. Symptoms and test results are compared with those of six other published cases. Results of skin tests obtained with a fraction of sperma plasma separated by chromatography are indicating that the antigen is of protein nature with a molecular weight of about 14.100. A molecular weight between 12.400 and 24.00 has also been reported in the literature.


[Diagnosis and therapy of stenosis or obstruction of the intrahepatic bile ducts]. 15 patients are reviewed who had stenosis or obstruction at the hilus of the bile duct. Percutaneous transhepatic or endoscopic transduodenal cholangiography is indispensable for establishing the diagnosis before operation. Primary carcinomas involving the junction of the hepatic ducts may be regarded as resectable if proximal extension of the tumor is limited to below the second bifurcation of the intrahepatic bile ducts in both the right and left lobes and if the proper hepatic artery and portal vein are still free from tumor invasion. Our favourable operation procedures for resection or palliative decompression of the biliary tree are discussed.


[Operative mortality, mortality and late results in bronchial carcinoma]. 457 patients with a bronchial carcinoma of the years 1947--1962 are compared with 126 patients of the years 1969--1970 and set up in comparison to the whole clinical material of the years 1960--1975. In the period between 1969 and 1970 radical resections were performed in 20% more patients than in the first period (1947--1962). Older patients were in the last period (1970--1974) more frequent. 8% of all patients  in which lobectomy was performed and 5.5% of the patients in which pneumonectomy was performed were older than 70 years of age. Overall mortality has remained unchanged for exploratory thoracotomy (19%), palliative resection (17.5%) and "radical" resection (14%). For simple pneumonectomy the overall mortality went down from 19.2% (1960--1964) to 9.6% (1970-1974) and for simple lobectomy from 14.4% to 3.8%. The five-year survival rate after radical resections was 8% (first period 0%) and after simple resections 26% (26.2%). Patients with stage I epidermoid carcinoma had a five-year survival rate of 46%. Those with stage II (spread to ipsilateral hilar nodes) a five-year survival rate of 14.3% Patients with palliative resections and exploratory thoracotomy died within 16 months.


Mutational biosynthesis of butirosin analogs. I. Conversion of neamine analogs into butirosin analogs by mutants of Bacillus circulans. By N-methyl-N'-nitro-N-nitrosoguanidine treatment, neamine-negative mutants which required neamine for biosynthesis of butirosins were obtained from a butirosin-producing organism Bacillus circulans. These mutants also produced butirosins from paromamine and could be divided into two types I and II. Mutants of type I could not produce butirosins from 2-deoxystreptamine, whereas those of type II could. Two typical mutants MCRL 5003 (type I) and MCRL 5004 (type II) could produce butirosin analogs, 3', 4'-dideoxybutirosins, 6'-N-methylbutirosins, 3', 4'-dideoxy-6'-N-methylbutirosins and 3', 4'-dideoxy-6'-C-methyl-butirosins from neamine analogs, gentamine Cla, 6'-N-methylneamine, 6'-N-methylgentamine Cla and gentamine C2, respectively.


Mutational biosynthesis of butirosin analogs. II. 3', 4'-Dideoxy-6'-N-methylbutirosins, new semisynthetic aminoglycosides. A pair of new butirosin analogs was isolated from the fermentation broth obtained by cultivating a neamine-negative mutant of the butirosin-producing organism Bacillus circulans in the medium supplemented with 6'-N-methylgentamine C1a. These antibiotics were characterized and elucidated as 3', 4'-dideoxy-6'-N-methylbutirosins A and B (DMB-A & DMB-B), by chemical and spectroscopic studies. DMB-A and DMB-B exhibited broad-spectrum antibacterial activities with in vitro potency similar to or slightly less than that for the butirosin A, with the exception of strains of Pseudomonas aeruginosa and Serratia marcescens against which they exhibited activities equal to or slightly greater than that for butirosin A. As expected, they exhibited stronger activities against butirosin-resistant organisms which contain acetylating enzymes AAC(6')-I and AAC(6')-IV, and phosphorylating enzyme APH(3')-II. They were also active against some of the clinical isolates resistant to butirosins, dibekacin and/or gentamicin. The acute intravenous toxicity in mice of the DMB complex (B:70 APPROXIMATELY 80%) was somewhat less than that of the butirosin A.


Mutational biosynthesis of butirosin analogs. III. 6'-N-methylbutirosins and 3', 4'-dideoxy-6'-c-methylbutirosins, new semisynthetic aminoglycosides. Two pairs of butirosin analogs were isolated from the fermentation broths obtained by cultivating a neamine-negative mutant of the butirosin-producing organism Bacillus circulans in the medium supplemented with 6'-N-methylneamine and gentamine C2, respectively. These amtibiotics were characterized as 6'-N-mentylbutirosins A and B (NMB-A & NMB-B), and 3', 4'-dideoxy-6'-C-methylbutirosins A and B (DCB-A & DCB-B), respectively, by chemical and spectroscopic studies. NMB-A and NMB-B exhibited broad-spectrum antibacterial activities with in vitro potency similar to or slightly less than that for butirosin A, but with greater activities against butirosin-resistant strain which contains acetylating enzyme AAC-(6')-I. The activities of NMB-A and NMB-B against Pseudomonas aeruginosa strains were relatively weak. DCB-B had broad-spectrum activity, and exhibited greatly improved activity against butirosin-resistant organisms which contain acetylating enzymes AAC(6')-I and AAC(6')-IV, and phosphorylating enzyme APH(3')-II. These new butirosin analogs were also active against some of the clinical isolates resistant to butirosins, dibekacin and/or gentamicin.


Ribostamycin production by a mutant of butirosin producing bacteria. By the use of our improved colony selection technique, xylostasin and ribostamycin producing mutants were isolated from nitrosoguanidine treated Bacillus circulans B15M, a producer of butirosins A and B. Among these structurally related aminoglycosides, ribostamycin is the well-known product of a Steptomyces and has not been isolated as a bacterial metabolite. A selected mutant of strain 306, which produces xylostasin and ribostamycin, was futher mutagenized in expectation of getting an improved strain having the ability to accumulate a large amount of ribostamycin in the culture broth. One mutant, strain 451, derived from strain 306, produced ribostamycin free of xylostasin.


1- and 3-deamidino derivatives of dihydrostreptomycin and some 1-N-acyl derivatives. 1-Deamidino-, 3-deamidino- and 1,3-di(deamidino)dihydrostreptomycin (1, 2, 3) were prepared by treatment of dihydrostreptomycin (DHSM) with ammonia at 100 degrees C. The 3-guanidino group of DHSM is suggested to be more important than the 1-guanidino group for the antibacterial activity of DHSM. 1-N-[(S)-4-Amino-2-hydroxybutyryl) and 1-N-[(S)-4-guanidino-2-hydroxybutyryl] derivatives (4, 6) of 1-deamidinodihydrostreptomycin were futher prepared.