Steady-state kinetics of low molecular weight (type-II) NADH dehydrogenase. (1) The steady-state kinetics of the NADH dehydrogenase activity of Type-II (low molecular weight) NADH dehydrogenase with the acceptors ferricyanide, cytochrome c and 2,6-dichloroindophenol are consistent with the simultaneous operation of an ordered and a ping-pong mechanism. Thus, depending on the acceptor concentration, the reduced enzyme is preferentially oxidized before or after NAD+ disociates from it. (2) The acceptors are able to oxidize the reduced enzyme and its NAD+ complex equally well. In contrast to the kinetics of the Type-I (high molecular weight) enzyme, double substrate inhibition is not found, implying that the site of oxidation of the reduced enzyme by acceptors and the NADH-binding site are remote. (3) With the indophenol, in the concentration range measured, the ordered mechanism is mainly operative. At infinite NADH and acceptor concentrations the rate constant of the reduction of enzyme by bound NADH is measured. (4) With ferricyanide and cytochrome c, in the concentration range measured, erroneous conclusions may be drawn from extrapolations owing to the fact that extrapolated lines in double-reciprocal plots of turnover number against acceptor concentration, at different NADH concentrations, intersect in the third quadrant. A method is described that allows the extrapolation of these data to zero acceptor concentrations. (5) The relation between activity and NADH concentration is sigmoidal (h = 2.0) with ferricyanide or cytochrome c as acceptor, but hyperbolic with 2,6-dichloroindophenol. The latter is also an inhibitor, competitive with respect to NADH. It is concluded that this two-electron acceptor, like ubiquinone, acts as an allosteric effector. (6) Type II is isolated from Type I without gross changes in tertiary structure, as judged by the unaltered rate constants of dissociation of NADH (k-1) and NAD+ (k4) and association of NADH (k1). (7) Type II differs from Type I in two respects, (a) The accessibility of the acceptors is greater by at least two orders of magnitude (k3). (b) The redox potential of the prosthetic group FMN is 120 mV less, as judged by a drop in the value of k2 by four orders of magnitude. It is suggested that one or more of the iron-sulphur proteins present in Type-I but lacking in Type-II dehydrogenase functions as an effector, regulating the redox potential of the FMN.


Electrochemical characterization of immobilized NAD+. Nicotinamide adenine dinucleotide (NAD+) has been covalently attached to alginic acid using carbodiimide coupling, thereby producing a macromolecular adduct of NAD, which can be rendered either soluble or insoluble by adjustment of pH. It was found that this NAD+-alginic acid complex was enzymatically active, and also that the oxidized form could be electrochemically reduced without loss in enzymatic activity. This NAD+ adduct has now also been polarographically characterized as to its two-step reduction waves, which are slightly shifted toward more cathodic potential as compared to free NAD+. When controlled electrolysis was conducted to reduce the bound NAD+ at the cathode, the NADH so formed by electrochemical action was found to be again oxidizable either enzymatically or electrochemically without loss in co-enzymatic function. The NADH adduct produced by electrochemical reduction of the NAD+ adduct has also been characterized by voltammetry.


Flash photolysis-electron spin resonance studies of photosystem I. A fast reduction of component of P-700+. A 300 mus decay component of ESR Signal I (P-700+) in chloroplasts is observed following a 10 mus actinic xenon flash. This transient is inhibited by treatments which block electron transfer from Photosystem II to Photosystem I (e.g. 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB), KCN and HgCl2). The fast transient reduction of P-700+ can be restored in the case of DCMU or DBMIB inhibition by addition of an electron donor couple (2,6-dichlorophenol indophenol (Cl2Ind)/ascorbate) which supplies electrons to cytochrome f. However, this donor couple is inefficient in restoring electron transport in chloroplasts which have been inhibited with the plastocyanin inactivators, KCN and HgCl2. Oxidation-reduction measurements reveal that the fast P-700+ reduction component reflects electron transfer from a component with Em = 375 +/- 10 mV (pH = 7.5). These data suggest the assignment of the 300-mus decay kinetics to electron transfer from cytochrome f (Fe2+) to P-700+, thus confirming the recent observations of Haehnel et al. (Z. Naturforsch. 26b, 1171-1174 (1971)).


Glycerokinase activity and lipolysis regulation in brown adipose tissue of cold acclimated rats. Glycerol release by brown adipocytes from constant cold adapted rats was not stimulated by norepinephrine. On the contrary, the release was stimulated in rats adapted to a nycthemeral fluctuatiing temperature from 5 degrees to 28 degrees C. Glycerokinase activity was greatly increased in brown adipose tissue by cold adptation ; there was no change in the liver. However this increased activity cannot entirely explain the lack of norepinephrine stimulation of glycerol release in the brown adipose tissue of cold adapted rats.


Association between macrophage activity and serum haptoglobin levels. Serum haptoglobin is elevated by treatment of mice with thioglycollate, B.C.G. or silica. Haptoglobin concentrations were followed for a two week period following treatment and were found to correlate with the known effect of the stimulants upon macrophage activity. The numbers and characteristics of peritoneal exudate cells present following intraperitoneal thioglycollate inoculation suggest a relationship between phagocytic activity and serum haptoglobin levels and support the idea that serum haptoglobin levels may be controlled by mediators released during the process of phagocytosis.


Temperature dependence of ESR spectra of spin-labelled met-hemoglobin. The maleimide spin-label, firmly fixed to the protein, was used to study conformation changes of various met-hemoglobin derivatives. The temperature dependence of the rotational correlation time shows a distinct conformation change in aquomet-hemoglobin at about 25 degrees C. The other met-hemoglobin derivatives studied (fluoro-, cyano-, aquomet-complexed with inositol hexaphosphate) and carbonmonoxy-hemoglobin exhibit no conformation changes in the temperature range from 0-50 degrees C.


[Mechanisms of transmission of excitation grnerated in the orbito-frontal cortex]. Experiments carried out on cats showed that interhemispheric generalization of strychnine spasmodic potentials from the orbito-frontal cortex were mainly brought about by the callosal pathway. In contrast to other projection and association zones of the cortex only the frontal portion of the corpus callosum (the rostrum) and not the whole corpus took part in transmitting strychnine commissures. The results of investigations carried out were consistent with the general conception of the principle of the determining dispatch station (DDS) in the nervous system integration activity. These results reveal that the appearance of the DDS induces secondary excitation foci which fully reproduce tha work regimen of the DDS. Switching of such "mirror" foci representing "the destination stations" from the DDS influence led to the elimination of the activity induced in them.


[Cyclic adenosine-3',5'-monophosphate content of the brain in conditions of extreme stimulation]. The content of brain cAMP was investigated under conditions of electric stimulation of immobilized rats which resulted in the development of reflex dystrophies of the internal organs. There was found a pronounced decrease in the amount of cAMP in the rat brain which developed 15 minutes after the beginning of extreme stimulation and remaained at the same low level for 3 hours after the electrical irritation. Reduction of the cAMP level was probably connected with noradrenaline and other biologically-active amine deficiency in the brain under conditions of extreme stimulation.


[Adrenaline-induced release from thrombocytes of beta-lipoprotein, alkaline phosphatase and 5'-nucleotidase]. Platelet aggregation induced by adrenaline is accompanied by release of beta-lipoprotein, alkaline phosphatase, 5'-nucleotidase and factor 3. Electrophoretic mobility of platelet alkaline phosphatase was the same as that of beta-lipoprotein. This suggest that beta-lipoprotein, alkaline phosphatase and 5'-nucleotidase are structural components of platelet compounds which possess factor 3 activity.


[Change in the mitotic activity of hepatocytes and resorption of necrotic areas in the formation of liver cirrhosis]. Cirrhosis of the liver was induced in rats weighing 120--140 g by CCl4 administration for a period of 2 months. Iodated oil, which caused emboly of the portal vein branches and foci of necrosis in the heptic tissue, was administered through the spleen to experimental and intact animals. The volume of necrotic foci, and also the mitotic actinity of hepatites were determined. Necrotic foci resolved more rapidly in cirrhosis of the liver. An increase of mitotic index of the second day after the administration of iodated oil was more significant in control rats.


[Effect of coniophage decomposition products on the biological activity of fibroblasts]. Cytochemical studies showed that destroyed coinophages, which phagocytized cytoxic and fibrogenic quartz dust, activated the synthetic function of rat fibroblasts when added to the culture medium. Inert talcum dust used under similar experimental conditions failed to produce any noticable changes in comparison with control studies.


Changes in the contractile response of arteires and veins from hypertensive rabbits to sympathetic nerve activity: assessment of some postsynaptic influences. Contractile responses to field stimulation of intramural nerves of arteries and veins taken from rabbits made hypertensive by partial constriction of the abdominal aorta have been related to the carotid artery pressure. The increase in contraction of cephalic and short saphenous veins with rise in carotid artery pressure can be accounted for by an increase in sensitivity of the alpha-adrenergic receptor. The neurogenic contraction of the ear artery increased with carotid artery pressure rise. Changes in some of the extraneuronal factors that influence transmitter distribution and disposition in the tunica media were examined. In hypertensive animals, the percentage of released adrenergic transmitter entering the vessel wall might be expected to decrease due to an increase in medial thickness. However, this percentage was not significantly altered in the ear artery probably due, in part, to a concomitant increase in medial permeability to the transmitter. Extraneuronal transmitter disposition factors, i.e. extraneuronal uptake, monoamine oxidase, and catechol-O-methyltransferase activity are directly related to the wet weight of the vessel wall. Thus, their contribution to transmitter disposition would be expected to increase with increase in vessel wall thickness and tend to reduce the response to sympathetic activity. As the contractile response increased in the hypertensive vessels despite such changes, the increase in effector cell mass and density of neuronal terminal plexus, shown previously to increase with hypertension, are more important than these other considerations.


Indomethacin-induced alterations in corticosteroid and prostaglandin release by isolated adrenocortical cells of the cat. 1 The effects of purported prostaglandin synthesis inhibitors on steroid and prostaglandin (E and F) release from trypsin-dispersed cat adrenocortical cells were investigated. 2 Low indomethacin concentrations potentiated adrenocorticotrophin (ACTH)-evoked prostaglandin and steroid release, whereas higher concentrations depressed both responses to ACTH. The steroidogenic response to exogenous prostaglandin E2 was not markedly altered over a wide range of indomethacin concentrations. 3 Indomethacin enhanced basal steroid release but did not enhance basal prostaglandin E or F release. 4 5,8,11,14-Eicosatetraynoic acid (ETA) elicited a concentration-dependent inhibition of ACTH-induced steroid release, but had little effect on prostaglandin E2-induced steroid release. A high concentration of ETA inhibited prostaglandin E and F release. 5 These data are discussed in relation to the concept that prostaglandins provide a critical link in ACTH-induced corticosteroidogenesis.


Van Buchem's disease (hyperostosis corticalis generalisata). The features of Van Buchem's disease are described, affecting six members of one family, spanning three generations of ages from 7 to 64 years.


A new stannous agent kit for labelling red blood cells with 99TCM and its clinical application. 99Tcm is chosen for labelling erythrocytes for vascular imaging to take advantage of the physical properties of the radionuclide. A new procedure is described which belongs to the category of pre-treatment methods using a kit. Stannous pyrophosphate is the specific reducing reagent. The technique is easier than already published methods: it consists of adding the above reagent to 2 ml. of heparinized blood, waiting five minutes and discarding the serum after centrifugation, then adding the radioactivity required as 99TcmO4. Labelling yield is superior to 95 per cent. The influence of technetium concentration of the pertechnetate solution on the labeling yield is evaluated and compensated by a slight adjustment of tin concentration when necessary. In vitro and in vivo stability studies show that 99Tcm-labelled red cells are suitable for determination of red cell volume, spleen and placental imaging, particularly in emergencies and in paediatrics.


Electrophysiological studies of normal and degenerating mouse neuromuscular junctions. A quantitative study of the functional changes occurring at end-plates of phrenicotomised mouse hemidiaphragms was made. Analysis of the frequency of spontaneous transmitter release revealed large scale deviations from a Poisson based process commencing 6-7 h post-phrenicotomy. Short term bursts of miniature end-plate potentials (minEPPs) lasting 0.5-1.0 sec frequently occurred and also long term bursts of minEPPs which lasted from several minutes to 0.5 h or more. Following the long term bursts there was regularly a lasting silence at the end-plates. MinEPP amplitudes were analysed. Probability analyses suggest that the minEPPs are often normally distributed amongst two or more populations. This relationship was maintained during degeneration of the end-plate. No significant differences between the distribution of mean minEPP amplitudes of degenerating end-plates and normal end-plates were found. This suggests that no change in the quantal unit of transmitter release occurs during degeneration of the end-plate.


NADH fluorescence, [K+]0 and oxygen consumption in cat cerebral cortex during direct cortical stimulation. NADH fluorescence, sagittal sinus blood flow and sinus hemoglobin saturation were monitored simultaneously during direct cortical stimulation of a wide area of the anterior and mid suprasylvian and marginal gyri. The area monitored flurorometrically was located within the area apparently drained by the sinus, so that the flurometric changes could be correlated with oxygen consumption changes calculated from the sinus flow and saturation values. The onset and peak values of calculated oxygen consumption and NADH fluorescence changes usually occurred within several seconds of one another and high, significant (r greater than 0.9 and P less than 0.01) correlations were found between the maximum changes in both parameters following stimulation. The relation of cortical [K+]0 changes to oxygen consumption changes was also explored; again the magnitude of [K+]0 changes and calculated oxygen consumption changes correlated well. The demonstrated agreement between fluorometric and direct (sinus cannulation) measurements of oxidative metabolism reinforces the interpretation of in situ cortical fluorescence changes as indicative of changes in oxygen consumption rate


Effects of kainic acid, a cyclic analogue of glutamic acid, on cyclic nucleotide accumulation in slices of rat cerebellum. The concentration of cyclic AMP (cAMP) and cyclic GMP (cGMP) in slices of rat cerebellum was increased markedly in the presence of 100 muM kainic acid, a cyclic analogue of glutamic acid. No acitvity was detected in other brain areas at various dosed or incubation times. Ksainic acid was significantly more potent than glutamic acid and other glutamate-like compounds. The increases in cAMP and cGMP could be differentially inhibited with theophylline or cocaine respectively, but both systems required calcium. Compounds similar in structure to kainic acid or glutamate did not block the effects of kainic acid.


Regulation of neuronal properties by afferent connections. I. Functional changes in snail neurons following section of presysnaptic nerve fibers. The left cerebral connective of snail ganglia containing afferent inputs to a particular neuron was sectioned. One week after the operation synaptic potentials were not obtained when the connective was stimulated, and they had not reappeared after 60 days. The current-voltage relationship of denervated cells showed a rectification in the hyperpolarizing direction, whereas that of control cells had an ohmic behavior. Depolarization or conductance changes produced by ACh or carbachol showed a supersensitivity in denervated cells, evidenced by a shift to the left of the dose-response curve and an increase of the maximal responses obtained. The reversal potential of ACh shifted from--22 mV in the control to--8 mV in denervated cells. Depolarization produced by serotonin was similar in control and denervated cells.


[Paget's disease of the breast in a male]. The case of a 78-year-old man with Paget's disease of the breast, described herein, appears to be only the 22nd histologically proven case of this rare condition; in over 100 years less than 50 cases have been reported. Since this patient had no underlying tumour or nodal involvement the operation consisted of total mastectomy with removal of axillary nodes only, rather than the classical radical mastectomy. All eczematoid lesions of the areola of the breast that persist for more than 2 weeks or are unilateral should be considered Paget's disease until proven otherwise by biopsy.


Oat cell carcinoma of the lung. Early treatment results of combination radiation therapy and chemotherapy. Since August 1974, 50 patients with histologically proven oat cell carcinoma of the lung were treated at Indiana University Hospitals with a combination of adriamycin, cyclophosphamide, Oncovin, BCG immunotherapy, and radiation therapy to the primary lung tumor and whole brain. Twenty-nine patients are eligible for a 6-month follow-up and are included in this report. All 29 patients responded to therapy; 12 patients (41%) had a minimum of 50% reduction in tumor mass and 17 patients (59%) had complete clinical remission of disease. Eight (38%) of the 29 patients have remained in complete remission from 6 to 14 months, and 90% of the patients survived a minimum of 6 months. This represents a considerable improvement in survival rates over what had been obtained at Indiana University Hospitals previously.


Postoperative radiation therapy for residual Wilm's tumor. Review of Group III patients in the National Wilm's Tumor Study. Fifty-eight patients with residual abdominal disease (Group III) are reviewed from National Wilm's Tumor Study "1. The totap III) are reviewed from National Wilm's Tumor Study No. 1. The total number of abdominal recurrences is relatively low (eight cases). Local irradiation of the renal fossa would seem to be sufficient treatment for disease believed to be limited to that region, including cases of local spillage. Whole abdominal irradiation is recommended for gross, diffuse peritoneal contamination.


Pulmonary function in survivors of Wilm's tumor. Patterns of impairment. Whole-lung irradiation in Wilms' Tumor patients has been given prophylactically and for treatment of metastasis. Thirty-three children with Wilm's tumor who have survived for 4 to 20 years from the time of diagnosis were avaiable for study. Whole-lung irradiation was given in doses from 1163 to 1370 rads (orthovoltage). Eighteen patients (Group N) received no irradiation to the lungs. The remaining 15 patients were divided according to whether pulmonary irradiation was given for metastatic disease (Group M--10 patients) or for prophylactic treatment (Group P--five patients). All but five patients in Group N received at least one course of actinomycin D. A routine set of pulmonary function tests was done to assess possible abnormalities of lung volume and the mechanics of breathing. Group N was normal. The findings in Group M suggested moderately reduced lung volumes and may have also reflected obstruction of the large airways and/or a limited expiratory effort. Group P had essentially normal lung volumes but also appeared to have obstruction of the larger airways. Abnormalities were generally not severe; only two patients, in Group M, were symptomatic. Excess irradiation, presence of metastatsis, additional lung irradiation, and pneumonitis may have contributed to morbidity.


Immune evaluation of lung cancer patients undergoing radiation therapy. Immunocompetence, as determined by skin reactivity to five microbial antigens, dinitrochlorobenzene (DNCB), and percent and absolute number of total T and B rosette-forming cells (RFC), was assessed before and up to 180 days after radiotherapy in 29 patients with carcinoma of the lung. Pretherapy evaluation showed a depressed response to microbial antigens (only 38% positive), to DNCB (50% positive), and lower level of T-RFC. Postradiotherapy determinations were further depressed. Patients with positive pretherapy responses to microbial antigens and DNCB and higher than median absolute numbers at total T-RFC had significantly longer survival. These data suggest that pretherapy immune evaluation is a good prognostic indicator. No prognostic significance was found in B-RFC evaluation. No correlation was seen between the stage of disease (locally advanced or with distant metastases) and the pretherapy immune response.


Urinary cholesterol. VIII. Its excretion in women with ovarian neoplasms. The urinary excretion of nonesterified cholesterol (NEC) has been investigated in 57 women with ovarian neoplasms and/or related nonneoplastic diseases. Twelve patients had benign tumors or lesions and 45 had malignant neoplasms of their ovaries. All patients with nonmalignant ovarian tumors or lesions had normal NEC excretion irrespective of the type of tumor or lesion or its degree of extension. In contrast, urinary NEC hyperexcretion occurred with the following frequencies in patients with active malignant ovarian neoplasms: 18 of 19 cystadenocarcinomas of the serous and/or mucinous types; one of one endometrioid carcinoma; four of four malignant granulosa cell tumors; two of two mixed malignant germ cell tumors; and one of one malignant mixed müllerian tumor. Single cases of clear cell carcinoma and of rhabdomyosarcoma had a normal NEC excretion. Urinary hyperexcretion of NEC was also found after surgery in two of seven surviving patients with apparently localized resectable disease according to their staging. It is possible that in these two patients NEC hyperexcretion was due to undetected foci of cancer (wrong staging), since neither omental and peritoneal biopsies, nor cytologic examination of peritoneal washings or free fluid were performed. A normal excretion of urinary NEC has been characteristic of 19 of 21 surviving patients treated by surgery and adjunctive therapy in whom we have performed follow-up NEC determinations. They were 16 of 18 cystadenocarcinomas malignant germ cell tumor. The 94% correlation between the presence of proven active ovarian carcinomas and urinary NEC hyperexcretion is significant. The clinical significance of this investigation is even greater when one considers that cystadenocarcinomas constitute more than 75% of all primary malignant ovarian tumors.


Radiation-induced intracranial malignant fibrous histiocytoma. An autopsy case of radiation-induced intracranial malignant fibrous histiocytoma (fibroxanthosarcoma) is reported. The tumor developed in the region of the sella turcica 11 years after high dose radiotherapy of a chromophobe adenoma of the pituitary. The tumor had infiltrated the base of the brain as well as the base of the skull. Metastases were not found. The tumor was composed of an admixture of bizarre fibroblasts, histiocytes and giant cells, xanthoma cells and siderophages, with a storiform fibrous stroma. This appears to be the first documented instance of a malignant fibrous histiocytoma occurring intracranially after local x-irradiation.


Signet-ring stromal tumor of ovary. Histochemical, light, and electron microscopic study. A peculiar primary ovarian neoplasm occurring in a 28-year-old woman is presented. The tumor cells showed extensive cytoplasmic vacuolation, resulting in a "signet-ring" appearance similar to what was described in Krukenberg tumors. The tumor was studied histochemically, as well as by light and electron microscopy. The cells appeared to be uniform. A moderately electron-dense flocculent material was present in the cytoplasmic vacuoles. similar material was seen in the intercellular matrix with some fibrillary structures suggesting a protein structure. This, coupled with the lack of any basement membrane material around the tumor cells, indicates a mesenchymal tumor that is probably derived from the multipotent ovarian stromal cell. The features that help to differentiate this tumor from other ovarian tumors with clear or vacuolated cells are discussed. Because of its gross and microscopic appearance, the tumor is expected to behave in a benign fashion.


Gastrinoma of duodenal G-cell origin.

Gastrinoma of duodenal G-cell origin. A 65-year-old man with hypergastrinemia associated with the Zollinger-Ellison syndrome was found to have a duodenal "carcinoid-islet cell tumor." Gastrin levels have remained normal for more than 1 year following total gastrectomy and removal of the duodenal tumor. Immunohistochemical studies for gastrin localization revealed positive staining of the tumor and of a population of nonneoplastic G-cells in the adjacent duodenal mucosa and Brunner's glands. These results support the hypothesis that gastrinomas may arise as primary tumors from duodenal G-cells rather than from ectopic pancreatic tissue. "Carcinoidislet cell tumors," like other tumors of APUD-cell origin, may express dual biochemical functions in the form of polypeptide hormone and/or amine secretion. Their content of specific hormonal products may be predicted on the basis of sensitive histochemical and immunohistochemical techniques.


Simultaneous occurrence of melanotic neuroectodermal tumor and brain heterotopia in the oropharynx. A case is described of heterotopic brain tissue with simultaneous occurrence of melanotic neuroectodermal tumor in the oropharynx of a 6-week-old infant. The melanotic neuroectodermal tumor was embedded within the heterotopic glial tissue. This coexistence leads us to speculate that a defect causing a pinching-off of both neural crest cap and medullary epithelium of neural tube might have taken place at, or before, the 25-30 somite stage. The displaced embryonic structures subsequently differentiated into glial tissue, choroid plexus, and melanotic neuroectodermal tumor. This observation may be interpreted as further support in favor of a neural crest origin of juvenile melanotic neuroectodermal tumor.


Familial glioblastoma with hepatic focal nodular hyperplasia. Two siblings succumbed to glioblastoma multiforme, associated with hepatic focal nodular hyperplasia and cafe-au-lait spots. One sibling had syndactyly and multiple colonic polyps, while the other had an angiomatous malformation in the brain. Their mother died of myasthenia gravis. The findings suggest a hamartomatous syndrome of malformations that predisposes to brain tumors.


Vesicocolic fistula due to "collision" between adenocarcinoma of the colon and transitional cell carcinoma of the urinary bladder. A patient with sigmoidocystic fistula due to "collision" between mucoid adenocarcinoma of the colon and transitional cell carcinoma of the urinary bladder is presented. A wide resection of the colonic tumor, along with the involved bladder segment, and a decompressive colostomy with bladder drainage by catheter, were performed. The clinical symptoms, diagnostic procedures, operative treatment, and prognosis are discussed. Although fistular formation is an indication of advanced carcinoma, wide resection of the tumor and the associated fistular along with the node-bearing area will result in a reasonable percentage of 5-year survivals.


Chemotherapy of invasive thymoma.

Chemotherapy of invasive thymoma. Despite conventional therapy, including surgery and radiation, 5-year survival for invasive thymoma remains poor. Chemotherapeutic agents, including cisdiamminedichloroplatinum, prednisone, adriamycin and nitrogen mustard-vincristine-vinblastine-procarbazine, and bleomycin-adriamycin-CCNU-vincristine have produced short-term remissions. In view of the small number of cases seen in any one institution, a cooperative study to evaluate chemotherapeutic efficacy in malignant thymoma would seem worthwhile.


Synovial sarcoma with brain metastases. Report of a case responding to supervoltage irradiation and review of the literature. A 32-year-old man with clinically evident brain metastases from synovial sarcoma had a complete clinical remission after cerebral radiation. At autopsy, residual tumor was found in the brain.


Hypothalamus and cytodifferentiation of the foetal pituitary gland. Study in vivo. To investigate whether the hypothalamus is involved in the cytodifferentiation of the anterior pituitary gland, rat foetuses were encephalectomized in utero on day 16 of pregnancy. Pituitary sections from encephalectomized and normal littermate foetuses were studied on day 21 with the immunofluorescence technique using antibodies anti alpha-MSH, anti beta-MSH, anti alpha-(17-39) ACTH and anti beta-(1-24) ACTH. On day 16, only the anti beta-MSH revealed a few cells in the pars distalis but not in the pars intermedia. On the other hand, on day 21, the pituitary cells reacting with antibodies anti alpha-MSH, anti beta-MSH and anti alpha-(17-39) ACTH were as numerous in the encephalectomized foetuses as in the normal littermate foetuses. The cells revealed with the antibody anti beta-(1-24) ACTH were less numerous and less fluorescent in the pars distalis and intermedia of the hypophysis of the encephalectomized foetuses. On day 21, the adrenals of the encephalectomized foetuses were atrophied in comparison with those of the normal littermate foetuses but they were larger than on day 16. These data suggest that the cytodifferentiation of the corticotroph and melanotroph cells of the hypophysis occurs without the influence of the hypothalamus which is necessary for the normal release of ACTH.


Fine structure of taste buds in the barbel of the catfish, Ictalurus punctatus. Taste buds occur in the epithelium of the catfish barbel along its entire length. Two major cell types, light and dark cells, occupy the upper two-thirds of the taste bud. A third cell type, the basal cell, lies on the basal lamina and is essentially separated from the light and dark cells by a plexus of unmyelinated nerve fibers. The dark cells have branching processes, both apically and basally whereas the light cells have a single apical process and many basal processes. The apical processes of dark cells contain secretory granules, while the apical processes of light cells contain an abundant agranular endoplasmic reticulum. Light cell nuclei contain bundles of 10 nm filaments, often arranged in the shape of a cup or ring, but nucleoli are rarely seen. It is suggested that this morphology indicates a low degree of RNA synthesis by light cells. The basal cells contain large numbers of vesicles, about 60 nm in diameter, which are sometimes seen in clumps in relation to an adjacent nerve fiber in a configuration resembling a synapse. Curiously, although basal cells present a large surface to the basal lamina, there are no hemidesmosomes. This suggests that the basal cell does not originate from the epidermis.


Specific binding of the type C viral core protein p12 with purified viral RNA. The major viral phosphoproteins (p12) of the Rauscher murine leukemia virus (R-MuLV) and the simian sarcoma-associated virus (SSAV) bind in vitro to their homologous 70S and 35S viral RNAs. Using purified 32P-labeled RNA and 125I-labeled p12 protein, complexes that are stabilized by formaldehyde-cross-linking can be readily detected after velocity gradient centrifugation. The in vitro reconstructed ribonucleoprotein complexes are seen only with p12 proteins incubated with viral RNAs isolated from the same type C viruses; no such complexes form with heterologous protein-RNA mixtures. Homologous but not heterologous p12 molecules compete with radiolabeled p12 protein for the specific viral RNA binding sites. The competition assay permits the detection of 10 ng of viral p12 protein. The major internal protein of type C viruses (p30) does not bind to viral RNA using identical assay conditions. From the specific activities of the radiolabeled components and also by equilibrium sedimentation analysis, we estimate that fewer than 15 molecules of p12 protein bind to each molecule of viral RNA. Both the specificity and stoichiometry of the p12-RNA interactions suggest that these RNA tumor virus proteins have a regulatory role in cells.


Accumulation of cystine auxotrophic thymocytes accompanying type C viral leukemogenesis in the mouse. Certain continuous lymphoid and myeloid tumor cell lines of rodent origin are unable to grow in tissue culture in the absence of pre-formed L-cystine (CYS). In contrast, three NZB murine lymphoid cell lines obtained from NZB mice free of hematopoietic neoplasm can grow as well in cystine-deficient media containing L-cystathionine (CSN), the immediate precursor of CYS in the biosynthetic pathway, as in cystine sufficient medium. The former class of cells is, therefore, CYS auxotrophs (CYS-) and the latter CYS prototrophs (CYS+). Compared to CYS+ cells, the CYS- lines appear to be relatively deficient in the enzyme cystathionase, which catalyzes the cleavage of CSN to CYS and alpha-ketobutyrate. Using protein synthetic capacity as a criterion, normal thymocytes from mixed-bred Swiss mice behave like CYS prototrophs, while those from littermates bearing Moloney type C virus-induced thymic tumors behave like CYS auxotrophs. The former are also characterized by substantially higher levels of cystathionase than the latter. Extracts of thymocytes from tumor-free AKR mouse thymus are also characterized by higher levels of cystathionase activity than extracts of spontaneous AKR thymomas. Exogenous in vitro type C virus infection of a CYS+ cell results in vigorous virus production but no concomitant reduction in cystathionase activity. Thus viral replication alone in any random lymphoid cell is not sufficient to alter the enzyme level. The data therefore suggests that CYS auxotrophy may closely accompany neoplastic transformation of certain hematopoietic cells in vivo, including that induced by certain "thymic" type C viruses.


Release of low density lipoprotein from its cell surface receptor by sulfated glycosaminoglycans. The sulfated glycosaminoglycan, heparin, was found to release 125I-labeled low density lipoprotein (125I-LDL) from its receptor site on the surface of normal human fibroblasts. Measurement of the amount of 125I-LDL released by heparin permitted the resolution of the total cellular uptake of 125I-LDL at 37 degrees C into two components: first, an initial rapid, high affinity binding of the lipoprotein to the surface receptor, from which the 125I-LDL could be released by heparin, and second, a slower process attributable to an endocytosis of the receptor-bound lipoprotein, which rendered it resistant to heparin release. At 4 degrees C the amount of heparin-releasable 125I-LDL was similar to that at 37 degrees C, but interiorization of the lipoprotein did not occur at the lower temperature. The physiologic importance of the cell surface LDL receptor was emphasized by the finding that mutant fibroblasts from a subject with homozygous Familial Hypercholesterolemia, which lack the ability to take up 125I-LDL at 37 degrees C, did not show cell surface binding of 125I-LDL, as measured by heparin release, at either 4 degrees C or 37 degrees C. Although heparin released 125I-LDL from its binding site, it did not release 3H-concanavalin A from its surface receptor, and conversely, alpha-methyl-D-mannopyranoside, which released 3H-concanavalin A, did not release surface-bound 125I-LDL. When added to the culture medium simultaneously with LDL, heparin prevented the binding of LDL to its receptor and hence prevented the LDL-mediated suppression of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity. The uptake of LDL by fibroblasts is proposed as a model of receptor-mediated adsorptive endocytosis of macromolecules in human cells.


Propagation of a segment of bacteriophage lamda-DNA in monkey cells after covalent linkage to a defective simian virus 40 genome. A 520 base pair DNA segment was excised from the bacteriophage lamda-genome by cleavage with the bacterial restriction endonuclease, endo R. Hindll. This segment was covalently joined in vitro to an 880 base pair simian virus 40 (SV40) DNA segment which contains the initation site for SV40 DNA replication. The latter segment was derived from the genome of a defective reiteration mutant of SV40 also by endo R. Hindlll cleavage. When the recombinant molecule, together with wild-type SV40 DNA as helper, was introduced into monkey cells by DNA infection, replication of the lamda-DNA sequences was observed, and hybrid genomes were encapsidated into progeny SV40 virions. The structure of the lamda-DNA segment after serial passage in monkey cells was examined by use of restriction endonucleases and electron microscopic heteroduplex analysis.


Topography of polyoma virus messenger RNA molecules. The different species of polyoma virus-spedific RNA molecules present in the cytoplasm of 3T6 cells 30 hr after viral infection have been characterized by molecular hybridization between nonradioactive polyadenlated RNA, fractionated by sedimentation through sucrose-formamide density gradients, and the 32P-labeled separated strands of restriction endonuclease fragments of polyoma DNA. Two relatively abundant RNA molecules, sedimenting at 16S and at 19S, transcribed from the L strand of the viral DNA, as well as a minor 20S species transcribed from the E strand of the DNA, were detected. The most abundant viral transcript, the 16S RNA molecule, was estimated to be complementary to the 22% of the L-strand DNA extending from 47 to 25 map units. The less abundant 19S L DNA strand transcript included all the sequences present in the 16S RNA and mapped between 68 and 25 map units. The minor 20S RNA molecule was tentatively identified as a transcript of the E-strand DNA from the entire early region of the polyoma genome. These three viral RNA molecules together exhaust greater than 95% of the coding capacity of the viral DNA. A small region of the DNA (4-5%), including the origin of DNA replication, does not appear to determine sequences present among the major stable species of vital mRNA.


Phenotypic mixing between N- and B-tropic murine leukemia viruses: infectious particles with dual sensitivity to Fv-1 restriction. In effort to understand how N or B tropism is determined in murine leukemia virus (MuLV) particles, we analyzed the MuLV produced after dual infection of mouse cells by N- and B-tropic MuLV. The progeny MuLV from such a mixed infection are sensitive to Fv-1 restriction in both N- and B-type cells, but are still highly infectious for mouse cells which do not exhibit Fv-1 restriction. This dual sensitivity to Fv-1 restriction is a phenotypic property of MuLV produced by mixedly infected cells, since individual virus clones derived from this MuLV are either N- or B-tropic. In further experiments, we superinfected murine sarcoma virus (MSV)-transformed cells with mixtures of N- and B-tropic MuLVs. The rescued MSV is restricted in its ability to transforms both N- and B-type cells. The results suggest that N- and B-tropic MuLVs specify different determinants, which are incorporated into virions along with the viral genome and which are the recognition sites for Fv-1 restriction. The presence of a given determinant in a virion renders the virus sensitive to restriction in cells of the opposite Fv-1 type.


The construction of viable nuclear-cytoplasmic hybrid cells by nuclear transplantation. Using the mouse L-cell line as a model system, a generalized approach is presented for nuclear transplantation in cultured cells resulting in the construction of cytoplasmic-nuclear hybrid cells. Techniques were developed for the preparation of cytoplast and karyoplasts having minimum contamination by parent whole cells. Sendai virusmediated fusion was performed in a manner which maximized the formation of the desired fusion products-cells having one cell equivalent of cytoplasm from one parent and a nucleus from a second parent. The viability of the fusion products was established by examination of photographic records of the developing cultures. Using these techniques, we found that nuclei could be introduced routinely into 10-30% of a cytoplast culture. From determinations of the increase in cell number with time, it was estimated that at least 30% of the reconstructed cells were capable of division. The approach was next applied to the formation of hybrid cells from L-cell cytoplasts and A9 cell karyoplasts. The A9 cell line is an azaguanine-resistant derivative of L cells. Thus any whole cells remaining in the culture of fused cells were readily eliminated by treatment with the purine analogue. The culture of remaining cytoplasmic-nuclear hybrid cells grew to confluence in the presence of azaguanine. The applicability of the approach to the construction of hybrid cells using parent lines from different organisms is briefly discussed.


Quantitation of elongating form A and B RNA polymerases in chick oviduct nuclei and effects of estradiol. The number of elongating form A and B RNA polymerases in chick oviduct nuclei was estimated by measuring incorportaion of 3H-UTP into 3' termini of nascent RNA chains in the presence of heparin to inhibit initiation, and quantitation labeled uridine released from these termini after alkaline hydrolysis. The method corrects for conversion of UMP to uridine (U) during manipulations and for production of 3' termini by ribonucleases and phosphatases. The results indicate that a large fraction of RNA polymerases elongating in vivo is retained in isolated nuclei: per diploid genome, approximately 1 x104 form B and 2 x103 form A enzymes are present. These levels are sufficient to maintain normal in vivo rates of mRNA and rRNA synthesis, but the average density of packing of polymerases on DNA is considerably less than the maximum density predicted by Miller and Bakken (1972), suggesting that initiation of polymerases of DNA is a limiting factor in the control of transcription. Rates of elongation of polymerases in vitro are severely impaired, indicative of a loss of elongation factors during nuclear isolation. After 6 hr of estradiol treatment in vivo, the level of form A enzymes in ovoduct nuclei increases to 5 x 103; little change in the number of form B enzymes is seen.


[Lysis of Rous sarcoma virus in the presence of anti-chicken fibroblast serum and complement]. Rous sarcoma virus produced by Chick embryo fibroblasts is inactivated by an antiserum prepared against uninfected fibroblasts in the presence, but not in the absence, of complement. This inactivation which demonstrates the presence of one or more antigens of the surface of the producer cell on the viral envelope, is due to virolysis. This is demonstrated by the release of the viral internal proteins and by the fact that the viral RNA becomes entirely degradable by RNase.


[Induction of endogenous C-type virus by 5-iododeoxyuridine and augmentation of this induction by polycyclic hydrocarbons]. When the MLg Mouse cells were treated with the chemical carcinogens, in the presence of microsomal enzymes and NADPH2 after 5-iododeoxyuridine (IUdR) treatment, the induction frequency of the endogenous C-type virus was increased by nearly 10-fold in comparison with the cultures treated with IUdR only. In the preliminary screening of several polycyclic hydrocarbons including carcinogenic and non-carcinogenic products, apparently there was no correlation between the carcinogenecity in vivo and the stimulation capacity of IUdR induced C-type virus activation in our test conditions.


[Prospective epidemiologic studies and the nature of the association between Epstein-Barr virus (EBV) and Burkitt's lymphoma (BL)]. The prospective study carried out by the International Agency for Research on Cancer on the relationships between E.B.V. infection and Burkitt's Lymphoma development as well as their partial malaria chemoprophylaxy proposals, are discussed as an example of studying the role of a virus and of co-factors in one human cancer.


[Absence of relationship between erythrocyte and viral antigen expressed on the surface of chick erythrocytes]. Anti-gs antisera were tested on embryo and adult Chick erythrocytes. Specific immunsera directed against Chick embryo or adult erythrocyte antigens have been tested on Hamster cell lines infected with the Rous virus. Results demonstrate that the erythrocyte and viral antigens are different.


[Increased transmission at electronic synapses and paroxysmal depolarization induced by cardiazol in the Mauthner cell]. Cardiazol induces in the Mauthner cell the paroxysmal depolarizing shifts which are characteristic of epilepsia at the neuronal level. The period of depolarizations is preceded and, later, accompanied by an increased transmission at the electrotonic synapses which are established upon this neuron by primary afferent vestibular fibers. Increased excitability of the chemical synapses occurs subsequently.


[Phosphorylase activity of human placenta perfused in vitro: effect of gonadotropins, dibutyryl cyclic AMP and prostaglandins]. The gonadotropic hormones stimulate the glycogen phosphorylase activity of human full-term placenta perfused in vitro. This effect is reproduced by dibutyryl cAMP and PG F2alpha and accompanied by an increased glucose liberation in the perfusion fluid.


Modification by prostaglandins E1 and E2, indomethacin, and arachidonic acid of the vasoconstrictor responses of the isolated perfused rabbit and rat mesenteric arteries to adrenergic stimuli. In isolated perfused rabbit mesenteric arteries, prostaglandin (PG) E1 and E2, 1-5NG/ML, did not alter the basal perfusion pressure, but reduced the vasoconstrictor responses to sympathetic nerve stimulation; the responses to injected norepinephrine were reduced by PGE1 and variably affected by PGE2. In contrast, in rat mesenteric arteries PGE1 and PGE2, 1-5 ng/ml, potentiated the vasoconstrictor responses to nerve stimulation and to injected norepinephrine. In rabbit mesenteric arteries, the inhibitor of PG synthesis, indomethacin, augmented the responses to sympathetic nerve stimulation and to injected norepinephrine, whereas in rat mesenteric arteries indomethacin inhibited the responses to both adrenergic stimuli. Arachidonic acid, a PG precursor, reduced the vasoconstrictor responses to sympathetic nerve stimulation and to injected norepinephrine in rabbit, whereas in rat, potentiation of the responses to adrenergic stimuli occurred. Since these effects of arachidonic acid were abolished by the simultaneous infusion of indomethacin, they appear to be mediated through conversion of arachidonic acid to PG. We conclude that prostaglandins modulate adrenergic transmission in mesenteric arteries and this effect is species dependent.


Influence of 5- and 6-hydroxydopamine on adrenergic transmission and nerve terminal morphology in the canine pulmonary vascular bed. We studied the effects of 5- and 6-hydroxydopamine on adrenergic neurotransmission, fluorescence histochemistry, and nerve terminal ultrastructure in the canine pulmonary vascular bed. Fluorescence histochemistry on stretched preparations and sections of intrapulmonary artery and vein demonstrated that these vessels are well supplied with adrenergic nerves electron microscopy revealed adrenergic terminals in the adventitia and outer third of the media in the artery, but only in the adventitia in the vein. Adrenergic terminals in artery and vein contained many small and a few large dense-core vesicles. At least 20% of the terminals in the artery contained many small agranular vesicles and a few large opaque vesicles; this suggests that they were of the cholinergic type; Such terminals were not found in intrapulmonary veins. Under conditions of controlled blood flow, stimulation of the sympathetic nerves to the lung and intralobar injection of norepinephrine increased pressure in the perfused lobar artery and small intrapulmonary vein in a stimulus-related manner. The rise in pressure in the lobar artery and vein in response to nerve stimulation was blocked after administration of either 5- and 6-hydroxydopamine; Neither agent modified the response of the pulmonary vascular bed to norepinephrine; In contrast, the rise in pressure in the lobar artery and vein in response to both norepinephrine and to nerve stimulation was blocked by phenoxybenzamine, an alpha-receptor blocking agent. The attenuated neurogenic vasoconstrictor response in dogs treated with 5- and 6-hydroxydopamine was associated with a marked decrease in intensity of fluorescence of the abundant adrenergic innervation in both intrapulmonary artery and vein, and with the appearance of an osmiophilic material in dense-core vesicles of adrenergic terminals in artery and vein. We believe that these data suggest that 5- and 6-hydroxydopamine interfere with adrenergic transmission in intrapulmonary vessels by depleting norepinephrine from adrenergic terminals. Furthermore, we conclude from hemodynamic, histochemical, and ultrastructural studies that vasomotor tone in the pulmonary vascular bed can be regulated by the sympathetic nervous system.


Myocardial tissue recruitment in the dog as determined by double tracer dilution method. The amount of tissue perfused, as determined from the difference in volume of distribution between a diffusible indicator (125I-antipyrine) and an intravascular indicator (131 I-albumin) was measured at different values of coronary flow, perfusion pressure, and vasomotor tone in the working left ventricle of an open-chest dog. Coronary pressure and flow were regulated independently from the systemic circulation and coronary vasomotor tone was reduced by dipyridamole. At each flow vasomotor tone was assessed by using as a reference the maximal vasodilation induced by arrest of flow. Measured tissue space was considered to be related to the capillary surface area available for tracer diffusion and therefore to the number of perfused capillaries per volume of muscle. A relationship between coronary blood flow and tissue volume was observed. It was found to be independent of vasomotor tone. Vasodilation was found to increase available exchanging capillary surface at a constant perfusion pressure.


An echocardiographic study of interventricular septal motion in the Wolff-Parkinson-White syndrome. Echocardiographic studies of interventricular septal motion were performed in 26 consecutive patients with the Wolff-Parkinson-White (WPW) syndrome and in ten normal subjects. All patients with types A or B pre-excitation were subclassified into groups I to IV on the basis of their electrocardiogram utilizing the method of Boineau and associates. In all 14 patients with type A (Group III or IV) pre-excitation, the motion of the interventricular septum and posterior left ventricular wall motion were normal. However, in 11 patients with type B (Group I) WPW an abnormal septal movement was noted. This was characterized in ten patients by an early systolic posterior motion, a subsequent anterior movement in mid systole, and the usual posterior septal motion beginning in late systole. In eight patients, including the one without early systolic posterior movement of the septum, the late systolic posterior movement was interrupted by a prominent septal notch. On e patient with type B (Group II) WPW was studied and exhibited normal septal and posterior wall motion. In one patient with a spontaneous change in the QRS complex from normal to a type B (Group I) WPW pattern, the septal motion was initially normal and abruptly changed following the first WPW beat. The onset of abnormal interventricular septal motion with type B pre-excitation QRS complexes strongly suggests that abnormal septal movement may be related to an altered sequence of ventricular depolarization during right ventricular pre-excitation.


Correlation of pathologic Q waves on the standard electrocardiogram and the epicardial electrogram of the human heart. To evaluate the relationship between abnormal Q waves on the standard ECG and localized ventricular excitation, unipolar epicardial electrograms were recorded over the left ventricle during aortocoronary bypass surgery in 36 patients. Of 20 without standard ECG Q waves, six had abnormal epicardial Q waves, three anterioly and three inferiorly. Of 16 patients with standard ECG Q waves, four had both precordial and anterior epicardial Q waves while seven had Q waves in leads III and aVF and inferior epicardial Q waves. Three of the 14 had Q waves in both precordial and inferior leads of the ECG but epicardial Q waves only from the antero-apical region in two and only from the inferior wall in one. Two patients with Q waves in both III and aVF had no epicardial Q waves. Thus, the standard electocardiogram underestimates epicardial Q waves. If Q waves are present in the standard ECG, they correlate with the presence, although not invariably the location of Q waves on the epicardial electrogram.


The His-Purkinje electrocardiogram in man: an initial assessment of its uses and limitations. A methodology is described for noninvasive recording of the electrical activity generated by the His-Purkinje system of man utilizing filtering, high amplification, and signal averaging. A waveform ranging between 1 and 10mu V was observed during the P-R segment. In many individuals, there was temporal overlap between the terminal P wave and the initial portion of the His-Purkinje waveform. In ten patients with long P-R intervals there was a strong correlation (r greater than 0.95) between the H-V time measured by electrode catheter and the duration of the His-Purkinje waveform. In two patients with atrial fibrillation the resultant His-Purkinje waveform was similar in morphology and duration to those observed in the ten patients. In each group H-V time was determined noninvasively and a waveform associated with electrical activation of the major portions of the His-Purkinje system was obtained.


Electrophysiological studies in patients with chronic recurrent ventricular tachycardia. Seventeen consecutive patients with chronic recurrent ventricular tachycardia (VT) were studied in an attempt to delineate the reproducibility and mechanism of this arrhythmia. Six patients had nonsustained and 11 had sustained VT. The following electrophysiological techniques were utilized in an attempt to reproduce VT: 1) rapid atrial and ventricular pacing (17 pts); 2) atrial extrastimulus technique (17 pts); 3) ventricular extrastimulus technique (17 pts); 4) V1V2V3 stimulation technique (5 pts); 5) ventricular pacing from two or more sites (5 pts). Ventricular tachycardia was induced in six of 11 (54%) patients with sustained VT. However, in four there was only a single induction and only in the remaining two patients could VT be repetitively induced. In the latter two patients ventricular tachycardia was induced with both atrial and ventricular stimulation. Ventricular tachycardia could not be induced in any patient with nonsustained VT, although three had spontaneous episodes of ventricular tachycardia during study. In conclusion, in the present series of patients with chronic recurrent VT, this rhythm could not be reproducibly induced in the majority of patients in the cardiac catheterization laboratory utilizing catheter stimulation techniques.


Observations on mechanisms of ventricular tachycardia in man. Fifty patients with sustained ventricular tachycardia were studied by endocavitary recordings and programmed electrical stimulation. Of 29 patients in whom tachycardia could be initiated, 18 had chronic coronary artery disease and eight had no detectable heart disease. Of 35 patients in whom the tachycardia could be terminated by premature ventricular stimuli, 21 had chronic coronary artery disease and eight had no detectable heart disease. Initiation of tachycardia was facilitated in 18 of 21 patients by pacing the ventricle at the slowest possible pacing rate. An inverse relation was found between the prematurity of the tachycardia-initiating premature beat and the interval between the premature beat and the first beat of tachycardia, a finding suggestive of a re-entry mechanism. The role of the specific conduction system in initiation and during tachycardia remains unknown. Stimulation site was found to affect initiation and termination of tachycardia and width of tachycardia zone.


Body-surface maps of heart potentials: tentative localization of pre-excited areas in forty-two Wolff-Parkinson-White patients. Heart potentials were recorded from the entire chest surface in 42 patients suffering from Wolff-Parkinson-White syndrome. We were able to identify six types of surface maps, according to the location of the potential maximum and minimum during the delta wave. For each of these types we suggested the most likely location of the pre-excited region around the A-V rings (types 1 to 5) or in the interventricular septum (type 6). In 13 patients belonging to Types 1, 2, 3, 5 and 6 our hypotheses were in agreement with intracardiac recordings, epicardial maps or surgical results obtained by others. Isopotential surface maps provide more information on the location of the pre-excited area than conventional ECGs, particularly when these exhibit intermediate features between Types A and B.


Serum lipoprotein profile in children from a biracial community: the Bogalusa Heart Study. Serum lipoprotein profiles in 3182 children, ages 5-14 years, were studied in a biracial community as part of the Bogalusa Heart Study to describe the early natural history of atherosclerosis. White and black children showed similar mean levels of beta-lipoproteins. Pre-beta-lipoprotein levels, however, were significantly higher in white shildren, while significantly higher levels of alpha-lipoprotein were found in black children. Girls had generally higher levels of beta- and pre-beta-lipoprotein and lower levels of alpha-lipoprotein than boys, although the differences were not significant at each age group. With age there was little change in alpha-lipoprotein levels, a significant increase in pre-beta-lipoprotein levels and a slight but significant decrease between 11 and 14 years in beta-lipoprotein levels. The correlation of alpha-lipoprotein was negative with beta-lipoprotein and, to a greater extent, with pre-beta-lipoprotein. The above inverse relationships were significantly greater in white children than in black children, suggesting differences in lipoprotein profiles in the two groups. Lipoprotein values from a total community study are now available for comparison with the currently recommended upper normal limits for lipoproteins. Since only a very small percentage of children could be considered as hyperlipoproteinemic by those specific levels in this community, we suggest that distributions and percentiles be used to evaluate children for hyperlipoproteinemia.


Cardioversion and "false positive" technetium-99m stannous pyrophosphate myocardial scintigrams. The present studies performed in experimental animals demonstrate that electrical direct current cardioversion can produce skeletal muscle damage and increased technetium-99m stannous pyrophosphate (99mTc-PYP) uptake; in experimental animals the electrically damaged skeletal muscle shows necrosis with extensive calcium deposition. In addition, the frequent administration of high energy cardioversion produces myocardial necrosis with calcium deposition, increased 99mTc-PYP myocardial uptake and a positive 99mTc-PYP myocardial scintigram. The data indicate that, if diagnostic 99mTc-PYP myocardial scintigraphy is contemplated after cardioversion, paddle placement should be slightly removed from the anteroposterior projection of the heart on the external chest wall to avoid possible subsequent confusion between increased myocardial and skeletal muscle uptake of 99mTc-PYP. If multiple high energy cardioversion episodes are necessary, myocardial necrosis resulting from electrical injury may occur and be responisble for increased myocardial uptake of 99mTc-PYP with a resultant positive 99mTc-PYP myocardial scintigram.


Paroxysmal supraventricular tachycardia: is the atrium a necessary link? Whether or not the atrium plays an essential role in initiating and/or sustaining atrioventricular (A-V) nodal re-entrant tachycardia was evaluated in eight patients. In all eight patients, the atrium could be rendered refractory to retrograde atrial echoes during the tachycardia without interrupting the arrhythmia. This was accomplished by introducing atrial premature depolarizations prior to the time the atrium would normally be retrogradely depolarized by atrial echoes. In one patient, two atrial premature depolarizations could be introduced, producing A-V dissociation, without terminating  the tachycardia. In another patient, the tachycardia could be initiated without an atrial echo. Our data suggest that most, if not all of the atrium is unnecessary for the initiation and maintenance of A-V nodal re-entrant supraventricular tachycardia.


Analysis of surgically-induced right bundle branch block pattern using intracardiac recording techniques. Using intracardiac recording techniques, His bundle (H) and right ventricular apical (RVA) electrograms were recorded in 16 patients with a postoperative electrocardiographic pattern of right bundle branch block (RBBB). Their ages ranged from 5 to 12 years (mean 6.9 years) at surgery and the follow-up period was 1 to 7 years (mean 2.7 years). All were asymptomatic and in sinus rhythm at the time of study. The P-A interval was normal in all and the A-H, H-V, and V-RVA intervals were prolonged in one, one, and six patients, respectively. The V-RNA interval was normal (less than or equal to 30 msec) in ten out of the 11 patients (91%) without associated left anterior hemiblock (LAH), indicating a physiologically intact main right bundle branch, and was abnormally lengthened (45-62 msec) in all five patients (100%) with associated LAH. These findings suggest that there are two subgroups of patients with surgicall-induced RBBB pattern and the measurement of the V-RVA interval in conjunction with the H-V interval may be of ultimate importance in understanding the long-term prognostic implication of surgically-induced RBBB pattern with or without LAH.


Earlier onset of QRS in anterior precordial ECG leads: precision of time interval measurements. Onset of QRS was compared between simultaneously recorded conventional ECG leads in 84 subjects with clinically normal hearts from a defined population sample. Mean onset of QRS was 6.4 msec earlier in lead V1 and 7.4 msec earlier in V2 than in lead II. These differences were statistically significant. The measuring system was adapted from drafting techniques and took into account variations in paper speed which occurred during recording. Interobserver differences equivalent to greater than 1 msec occurred in 3.9% of timeline measurements, but in in 38% of QRS onset measurements. The lower precision in measuring QRS onset may be attributed to baseline oscillations and to the relatively slow rate of voltage change at the onset of ventricular depolarization.


Treatment of a case of allergic asthma with a synthetic human ACTH preparation. The following case history documents the first patient treated with synthetic human ACTH. It was prescribed for an asthmatic child because of suspected allergy to both porcine ACTH and tetracosactrin. It proved both effective clinically and free of any adverse effects.


Purification and analysis of the purity of NADH. We developed an analytical reverse-phase high-performance liquid chromatographic procedure for rapid assessment of the purity of NADH. The method completely separates adenosine monophosphate and adenosine diphosphoribose from NADH. By use of this analytical technique we found that preparative chromatography on DEAE-cellulose gives NADH that is free from adenine nucleotides as well as other impurities that commonly are present in NADH. The absorbance ratio at 260 and 340 nm of the purified NADH in 1.8 mmol/liter ammonium carbonate is 2.261 +/- 0.002 (+/- 1 SD).


Structure of some aliphatic dicarboxylic acids found in the urine of an infant with congenital lactic acidosis. Gas chromatography/mass spectrometry was used to identify a series of acids in urine and serum from a child who died 26 h after birth in severe metabolic acidosis with high lactate excretion. cis-5-Decene-1, 10-dioic acid and cis-5-dodencene-1, 12-dioic acid were synthesized and used as references. The following acids were found: hexane-1,6-dioic acid, octane-1,8-dioic acid, decane-1,10-dioic acid, dodecane-1,12-dioic acid, cis-5-decene-1,10-dioic acid, cis-5-dodecen-1,12-dioic acid, cis-5-tetradecene-1,14-dioic acid, trans-3-decene-1,10-dioic acid, and trans-3-dodecene-1,12-dioic acid. The concentration of C6 to C14 acids in the patient's urine was 3.7 mol/mol of creatinine; it was less than 0.2 mol/mol of creatinine in eight normal newborns and approximately 0.1 mol/mol of creatinine in a case of fructose-1,6-biphosphatase deficiency with lactic acidosis. 5-cis-Dodecenedioic acid was present in highest concentration: 1 mol/mol of creatinine in urine and 61 mumol/liter in serum. We propose that impaired beta-oxidation, probably at the acyl-CoA-dehydrogenase step, resulted in the formation of the observed acids. The parents were consanguineous, and a sibling died with the same clinical picture, which suggests a genetic defect.


Correlation between estrogen receptor protein and carcinoembryonic antigen in normal and carcinomatous human breast tissue. We determined estrogen receptor protein and carcinoembryonic antigen in cytosols prepared from 189 human breast carcinoma tissues, 85 benign or normal breast biopsies, and 101 tissue specimens metastatic from breast carcinoma. Carcinoembryonic antigen was observed in 70% of the primary carcinomatous tissues, 15% of the benign or normal specimens, and 51% of the metastases. Ninety-six of the 189 primary carcinomatous specimens with increased concentrations of carcinoembryonic antigen were also positive for estrogen receptor protein, whereas 67 of the 72 benign or normal biopsies with low concentrations of carcinoembryonic antigen were also negative for estrogen receptor protein. All five fenign specimens with positive estrogen receptor protein and normal carcinoembryonic antigen concentrations were from fibroadenomas. The concordance between estrogen receptor protein and carcinoembryonic antigen in the primary carcinomatous tissue was 66%, in metastatic carcinoma 51%, and in benign and normal tissue 85%.


End-point parameter adjustment on a small desk-top programmable calculator for logit-log analysis of radioimmunoassay data. We describe an improved method of logit-log curve fitting, by adjusting end-point parameters in radioimmunoassay studies, for use with a small desk-top programmable calculator. Straight logit-log analyses are often deficient because of their high sensitivity to small errors in the end-point parametes B0 and NSB (the actual measured activity in the tubes). The literature suggests techniques for adjusting these end-point parameters, but they require too much computing time and programming space to be used with a desk-top programmable calculator. The extension to the logit-log model presented here is easily handled by the programmable calculator and provides a good estimate of the change required in B0 and NSB to obtain a better fit. The program requires 1.5 min to run on our desk-top programmable calculator, and has resulted in improved data analysis for all of the 11 types of radioimmunoassay studied.


Unsuitability of porcelain crucibles for dry ashing of biological tissues for calcium analysis. At temperatures required for complete release of calcium from beef liver by dry ashing, porcelain crucibles release significant amounts of calcium into the ash, which leads to erroneously high calcium values in the samples. Heated, empty porcelain crucibles do not show released calcium. Both fused silica and platinum crucibles fail to show any significant interaction with the sample. Samples dry-ashed in fused silica crucibles were compared to samples subjected to the hot trichloracetic acid-acetic acid extraction method, which gave results not significantly different if the tissue was first minced. However, calcium was incompletely extracted by the hot acid from whole segments of liver weighing about 0.2 g.


Primary hyperlipoproteinemia in xanthomatosis. Blood lipid values, clinical data and effects of therapy are reported on 74 patients with hyperlipidemia and xanthomatosis. A natural subdivision into two groups was observed on the basis of low density lipoprotein lipid values: one corresponding to Frederickson's type II, characterized by elevated low density lipoproteins, tendinous xanthomata, absence of eruptive xanthomata and a high incidence of cardiovascular diseases and the other resembling Frederickson's type III, with elevated very low density lipoproteins, eruptive xanthomata, xanthomata striata palmaria, elevated cholesterol/triglyceride ratios in the very low density lipoproteins and irregular appearance of floating beta lipoproteins. The latter group consisted of 32 patients in whom cardiovascular symptoms were relatively rare, despite mean cholesterol levels of 500 mg/dl.


Lack of cytochalasin E-induced superoxide release by polymorphonuclear leucocytes of patients with chronic granulomatous disease: a new diagnostic test. Reduction of exogenous cytochrome c was induced by the treatment of human polymorphonuclear leucocytes with cytochalasin E. The reduction was completely inhibited by superoxide, dismutase, indicating that superoxide anions were released from the cells. The leucocytes from two patients with chronic granulomatous disease did not show the release reaction that should have been brought on by the treatment. The cytochalasin-induced reduction of exogenous cytochrome c by polymorphonuclear leucocytes is so specific and sensitive that it seems to be useful for diagnosis of chronic granulomatous disease.


Maternal immunoglobulins and parainfluenza 3 virus inhibitors in the nasal and lachrymal secretions and serum of newborn lambs. Concentrations of IgG, IgM and IgA were measured in the serum, nasal secretions and lachrymal secretions of suckled newborn lambs. The major immunoglobulin constituent of precolostral serum was IgM. Maternal immunoglobulins, of which IgG was predominant, reached peak values on day 1 of life and then declined over the next 3 weeks. Half lives were calculated as: IgG, 13-7 days; IgM, 4-1 days; and IgA 1-8 days. No immunoglobulin was detectable in nasal or lachrymal secretions prior to sucking but IgG was present in all samples of these secretions obtained approximately 24 hr after first sucking. IgG was present in nasal washings from suckled lambs, reared either naturally or on immunoglobulin-free milk substitute and levels declined as the lambs grew older. IgM and IgA did not appear consistently in the secretions until lambs were 2-3 weeks old. It was concluded, therefore, that colostral IgG reaches the nasal and lachrymal secretions of the newborn lamb. However, because the ewes in this experiment had only low serum titres, no maternal antibody to parainfluenza 3 virus (PI3) was detected in the nasal secretions of the lambs, although non-specific inhibitors were present. It is suggested, however, that low levels of maternal antibody in the secretions may play a valuable role in preventing respiratory virus infections of young ruminants before active local production of IgA and IgM begins at 2 to 3 weeks of age.


The stimulation of lymphocytes by chondrocytes in mixed cultures. The stimulation of lymphocytes by allogeneic and syngeneic cells other than lymphocytes has been shown to result in poor or no stimulation. We have been able to demonstrate that lymphocytes and chondrocytes mixed in culture both allogeneically and syngeneically has resulted in siginificant lymphocyte stimulation. The inference from this data suggest the presence of histocompatibility antigen as well as tissue-specific antigen on the surface of chondrocytes.


Transmission of auto-immune haemolytic anaemia and murine leukaemia virus in NZB-BALB/c hybrid mice. When mated to normal BALB/c partners, male and female NZB mice transmitted auto-immune haemolytic anaemia to three generations of their hybrid progeny. Red cell auto-antibodies (positive Coombs tests) were detected, on average, 11 months later in the F1 hybrids than in the parental strain, and the course of the disease was protracted. In explicably, the auto-immune reactions then appeared progressively earlier in successive generations of both croses. The Coombs reactions of the F1 and F2 hybrids were often weak and inconsistent, while those of F3 offspring showed the strong and stable picture typical of NZB mice. The incidence of auto-immune disease in each generation, although similar in the reciprocal crosses, indicated that the pattern of inheritance was very complex. The hybrids did not develop the lymphoid type B reticulum cell neoplasia which characterizes auto-immune NZB mice. Instead, and irrespective of Coombs status, they had lymphocytic leukaemias, lung adenomas, hepatomas and type A reticulum cell neoplasms of the liver. Murine leukaemia virus was identified electronmicroscopically in F1 embryos, and in all the adults examined. It was also isolated from leukaemic spleen cells passaged briefly in vivo, and from malignant hepatic (reticulum) cells maintained in vitro. These isolated were leukaemogenic in newborn BALB/c, NZB, and F1 hybrid recipients, but did not induce or accelerate positive Coombs reactions. Only a small proportion of the hybrids had significant glomerulonephritis, and overt kidney disease was minimal. The lesions were not confined to Coombs-positive mice. Possible links between auto-immunity, malignancy, and virus infection in NZB mice are discussed in the light of these results.


Properties of blast cells in 'control' cultures of human blood lymphocytes. Preliminary note. Ficoll-Hypaque separated normal human peripheral blood lymphocytes undergo appreciable blastogenesis in RPMI 1640-pooled human serum or - autologous plasma. During the 1st week, granulated blasts with peroixdase activity and ultrastructural properties of granulocytes precursors appear. These cells, which appear to be relateed to CFC (colony-forming cells), contribute to the mitotic population in these unstimulated "control" cultures prior to 6 days.


The effects of cyclic AMP on leucocyte inhibitory factor (LIF) production and on the inhibition of leucocyte migration. The effect of drugs known to increase intracellular levels of cyllic AMP were studied in the leucocyte migration ihibition system. It was found that cyclic AMP, dibutyryl cyclic AMP, theophyline, and prostaglandins E1 and E2 inhibited the production of leucocyte inhibiting factor by HA pulsed lymphocytes Inhibition only occured when the drugs were present during or after the PHA pulse. In addition it was found that these drugs enhanced the migration of polymorphonuclear leucocytes (PMN), in this system. Electrophoretic mobility of PMN cells was not altered by these drug indicating that the effect is not due to changes in membrane charge. However, granulocyte adhesion was reduced in the presence of these drug suggesting that adhesion is of primary importance in the migration of polymorphonuclear leucocytes out of capillary tubes. The findings show that cyclic AMP is important in modulating both cell-mediated and inflammatory responses.


Immunological studies of human placentae. Lectin binding to villous stroma and to trophoblastic basement membrane. It has been demonstrated immunohistologically using cryostat sections that the lectins PHA and Con A bind to human placentae. Both lectins stain trophoblastic basement membrane (TBM), perivascular tissue and stroma. No staining of trophoblasts was observed. It has been shown by blocking experiments that the lectin binding sites on placental TBM are glycoproteins, whereas experiments involving pretreatment of placental sections with anti-collagen antisera or highly purified bacterial collagenase have indicated that lectin binding to stromal and perivascular structures is collagen-associated. The possible relation of TBM lectin-binding sites to immune response gene products is briefly discussed.


Lithium treatment during alcoholic withdrawal. The efficacy of lithium carbonate was studied in 18 chronic alcoholic male patients in withdrawal. In mild alcoholic withdrawal, oral lithium carbonate, 0.3 gm every 8 hr, diminishes subjective symptoms of withdrawal and normalizes performance on a motor tracking task. Patients who start lithium while drinking ethanol improve most probably because it takes longer than 3 days for lithium concentrations in the blood to plateau. Lithium does not importantly alter patterns of catecholamine excretion, blood pressures, heart rate, serum cyclic-adenosine monophosphate (AMP), serum dopamine beta-hydroxylase (DBH), sleep pattern, or tremor amplitude during withdrawal.


Reversible resistance to the renal action of parathyroid hormone in man. 1. Normal subjects showed a highly reproducible, rapid increase in plasma adenosine 3':5'-cyclic monophosphate (cyclic AMP) after an intravenous injection of 200 MRC units of highly purified bovine parathyroid hormone. 2. No significant increase in plasma cyclic AMP was observed after administration of bovine parathyroid hormone to patients with severe chronic renal failure. 3. Even when renal function was not impaired, some patients with primary hyperparathyroidism, who had high concentrations of endogenous parathyroid hormone, showed resistance to bovine parathyroid hormone and when this was injected intravenously it caused only a small increase in plasma cyclic AMP. This resistance was reversible since there was marked improvement in the response after parathyroidectomy, when endogenous parathyroid hormone concentration had fallen. 4. It was possible to reproduce this resistance to the hormone by intravenous infusion of bovine parathyroid hormone into normal subjects. When the hormone (1000 MRC units) was infused over 2 h, after an initial increase there was a progressive decline in plasma cyclic AMP concentration and a fall in urinary cyclic AMP excretion. The response to a standard test stimulus (200 MRC units of bovine parathyroid hormone given as a rapid intravenous injection) was examined at intervals after 1000 units of bovine parathyroid hormone had been infused. Initially, the response was severely impaired; at 4 h, partial recovery had occurred and, 24 h after the infusion, recovery of the response was complete. The resistance was therefore reversible. Infusion of the amino-terminal peptide, fragment 1-34, gave the same effect as infusion of intact hormone. Region-specific assays for the hormone were used to show that the concentration of immuno-assayable hormone remained high during the infusions. 5. The mechanism of this reversible resistance to parathyroid hormone remains to be elucidated; it seems unlikely that circulating hormone fragments could account for the prolonged impairment in the responsiveness to the intact hormone. It is possible that alteration in the formation, intracellular degradation or, perhaps, release of cyclic AMP from the cells, is the cause. Changes in the characteristics of the hormone receptor sites might also explain the phenomenon.


Stimulation-induced depletion of vesicles, fatigue of transmission and recovery processes at a vertebrate central synapse. The Mauthner fiber giant fiber synapses of the hatchetfish are chemically transmitting axo-axionic synapses in the medulla. Tetanic stimulation at room temperature depletes the presynaptic Mauthner terminal of vesicles and leads to the appearance of large numbers of irregular membraneous compartments in the terminal. Stimulation during cooling to 12 degrees C depletes the terminal of vesicles and greatly increases the external surface, which forms large whorls of invaginating double membranes. Many coated vesicles are attached to the surface and the invaginating whorls. It is concluded that vesicles are discharged by exocytosis and fusion of their membrane with the external surface, and that at room temperature, membrane is reinternalized by coated vesicles and formed into irregular compartments. In completion of the cycle, these compartments disappear, and the vesicle population recovers over an hour or two of rest. When the Mauthner fibers are stimulated at low rates, the p.s.p.'s in the giant fibers are large and suprathreshold. Minature p.s.p.'s are generated spontaneously or can be evoked by subthreshold depolarization or tetanic stimulation of the Mauthner fiber. Stimulation of the Mauthner fibers at gradually increasing frequencies depresses p.s.p. amplitude to or below the level of miniature p.s.p.'s, but no failures are observed. Small p.s.p.'s without failures suggest that the quantum number remains high but that quantal size is greatly reduced, either by partial filling, as is supported by the morphological observation of vesicle depletion, or by desensitization. When stimulation is stopped, recovery of p.s.p. amplitude occurs in 1 or 2 seconds, but if tetanic stimulation is resumed immediately, p.s.p. amplitude decreases again and much more rapidly than in the initial rundown. This result suggests that depression of p.s.p. amplitude is not due to desensitization and leaves partial filling as the most likely explanation of small quanta. Calculated quantal size following a tetanus recovers in 200-500 ms, which probably largely reflects the time for filling since enough vesicles can be supplied to prevent failures with much shorter intervals between stimuli. Because quantal size appears to decrease gradually as stimulation frequency increases, it appears that release of vesicles can interrupt filling, leading to the conclusion that filling and release sites are very close together. This conlusion is consistent with other data in the literature obtained by different techniques.


Synaptic organization and acetylcholine sensitivity of multiply innervated autonomic ganglion cells. The principal cells of the mudpuppy cardiac ganglion receive synapses from three sources: vagal axons, interneurons and axon collaterals from other principal cells. The simplicity of the structural organization and the visual clarity in the living preparation provide favorable conditions for examining the function of these synapses and how different classes of synapses on the same cell influence its function. We have studied the sensitivity of the principal cells to iontophoretically applied acetylcholine--the transmitter at synapses made by the vagal axons and by postganglionic axon collaterals from other principal cells. In normal ganglia, the ACh sensitivity on the cell surface is highest at the region of synapses. Partial denervation, produced by severing the vagus nerves, results in an increased ACh sensitivity in nonsynaptic areas but does not appear to affect synaptic transmission at the remaining synapses.


The structure and permeability of isolated hepatocyte gap junctions. The ultrastructure of the gap junction may be visualized in both in situ and isolated preparations by using a variety of electron microscope techniques. The junction is composed of a lattice of subunits, called connexons, which show variable degrees of packing into a hexagonal lattice depending on a variety of poorly understood conditions. In general, it appears that more uncoupled and "dead" the junction, the more regular and condensed the hexagonal lattice becomes. It is not yet known whether these are "postmortem" changes or physiologically active and reversible changes involved in regulation of junctional permeability. Using a variety of techniques, it can be seen that the connexon extends completely across the junctional membranes, from the cytoplasmic surface of one cell to the cytoplasmic surface of the other, spanning the 2-nm "gap" between the apposed junctional membranes. Thus it is possible to implicate the connexon as a permeability channel from cytoplasm to cytoplasm, although the hydrophilic pore through the center of the connexon has not yet been demonstrated to span the full junction thickness. X-ray diffraction experiments support these conclusions, and the excellent correlation between the electron microscope and X-ray diffraction data lends great confidence to the interpretations of gap junction structure presented thus far. These data are summarized in the drawing in Figure 17. This is a scale drawing of two connexons, each of which is imagined to be composed of a dimer of hexamers. It must be emphasized that as yet there is no direct evidence for a sixfold symmetry within the connexon. Of special interest now are the types of protein-protein interactions that hold the two halves of the connexon together across the 2-nm gap and the lateral interactions between the connexons at the level of the lipid bilayers.


Modification and regeneration of synaptic connections in cultured leech ganglia. Segmental ganglia of the central nervous system of the leech were maintained in culture medium outside the animal for several weeks in order to study the properties of synapses and regeneration by identified sensory and motor nerve cells. A variety of preparations were used, including single ganglia, chains of ganglia and ganglia connected to the areas of skin and muscle that they normally innervate in the animal. (1) For up to 10 weeks after removal from the animal, resting and action potentials recorded from sensory and motor neurons resembled those seen in normal ganglia. The same individual cell in a cultured ganglion could be recorded from with intracellular electrodes on a second occasion after an interval of a few days. (2) Sensory cells, identified as touch, pressure or nociceptive according to their morphology and electrical properties, continued to respond selectively to stimuli of the appropriate modality applied to their receptive fields in the skin; action potentials in motor cells caused contractions in the appropriate muscles. Culture of ganglia for more than 3 weeks caused the disappearance of synaptic potentials and a loss of transparency in ganglia. (3) Certain chemically mediated synaptic interactions between sensory and motor nerve cells became markedly changed in cultured ganglia. These changes appeared over the first 3 weeks and consisted of abnormally large excitatory and inhibitory synaptic potentials. The changes in synaptic transmission observed in culture were in many respects similar to those occurring in ganglia maintained within an animal after lesions have been made in the nervous system (Jansen et al. 1974). (4) The morphological appearances of sensory cells were compared in cultured and normal ganglia after injection of horseradish peroxidase. In cultured ganglia, the branching pattern appeared normal, but varicosities became more conspicuous. (5) When connectives linking cultured ganglia were crushed or cut, regeneration occurred. By 7 days, impulses propagated through the regenerated fibers and evoked synaptic potentials on cells within the next ganglion. The course taken by regenerating axons was observed in cells injected with horseradish peroxidase. The results again resembled those seen in animals with similar lesions. (6) The cultured ganglia provide preparations in which it is possible to analyze the mechanisms that underlie long-term changes similar to those seen in the leech central nervous system in situ.


Transmission of signals from photoreceptors to ganglion cells in the eye of the turtle. Synaptic transfer between receptors and ganglion cells was studied in the retina of the turtle. In the normal operation of this pathway, signals are relayed across two or more chemical synapses. Previous work indicates that the receptors give graded hyperpolarization to light, and that the bipolar cells also respond with graded potential changes; the ganglion cells give impulses. Injection of weak electrical currents into a single receptor provides an alternative means of activating the pathway. We have used this technique to examine (1) the role of the receptors' hyperpolarization in signal transmission and (2) the sensitivity and temporal properties of the pathway. The response of a ganglion cell evoked by light on the retina can be duplicated by electrically hyperpolarizing a receptor and antagonized by electrical depolarization. This indicates that the hyperpolarization is responsible for regulating the flow of information from the receptors to the second-order retinal cells. The sensitivity of the synaptic path from cones to ganglion cells, determined from electrical stimulation of single receptors, was found to be sufficient to permit detection of about 100 photoisomerizations. Using the same technique, the kinetics of transfer were studied and found to exhibit a delay or integrating process and also a slow differentiation that blocks steady-state transmission. For the path from red-sensitive cones to ganglion cells, the apparent time constant of the delay process was of the order of 75 ms and that of the differentiation, about 100 ms. In the path from rods to ganglion cells, the differentiation was several times slower.


Reappraisal of the role of the diabetic state in coronary artery disease. Clinical and coronary arteriographic findings were evaluated in patients with angina pectoris who were considered not to have diabetes mellitus or to have chemical or clinical diabetes. Each of the three groups consisted of 100 consecutive referred patients. Neither the age of the patients nor duration of symptoms differed significantly among the groups. Hypertension, gout, and peripheral vascular disease were more frequent in the patients with clinical diabetes. There was no difference in serum cholesterol concentration among the groups, but plasma triglyceride levels and the frequency of type 4 hyperlipoproteinemia were significantly higher (p less than 0.01) in the chemical and clinical diabetic groups than in the nondiabetic patients. Coronary arteriographic observations indicated that the severity of the coronary arterial disease was greater in both diabetic groups than in nondiabetic patients. The difference in the coronary scores among the three groups of patients interacts to some extent with the triglyceride level, since a high score in the diabetic groups was noted only in the presence of an elevated tryglyceride concentration. The results indicate that the increased severity of coronary arterial disease in diabetic patients is not attributable to age, duration of symptoms, hypertension, type -4 hyperlipoproteinemia, or apparent severity of the glucose intolerance.


Another presentation of fibrosing alveolitis and alveolar cell carcinoma. A 73-year-old woman had dyspnea associated with gradually increasing pulmonary collapse. Despite intensive investigation, the diagnosis was only reached at autopsy 18 months later, when histologic examination of the lungs showed changes of both fibrosing alveolitis and alveolar cell carcinoma.


Onset of sarcoidosis with left ventricular failure and multisystem involvement. A patient presented with acute left ventricular failure, then developed cranial nerve palsies, myopathy, fever and hyperuricemia. Sarcoidosis was not diagnosed until the pulmonary congestion cleared, revealing hilar adenopathy and residual mottling. Thereafter, uveitis was noted and scalene and muscle biopsies confirmed the diagnosis.


Characterization of a nosocomially significant, multiple drug-resistant strain of Serratia marcescens. A multiple drug-resistant strain of Serratia marcescens (bacteriocin type 18) was isolated from three clinical patients. The isolates were found to carry a conjugally nontransferable, nonmobilizeable resistance plasmid (R-plasmid) with resistance-(r)-determinants against ten antimicrobial drugs: ampicillin, carbenicillin, chloramphenicol, gentamicin, kanamycin, neomycin, streptomycin, tobramycin, triple sulfonamides, cotrimoxazole, and--possibly--nalidixic acid, as determined with exposure to 'curing' agents (ethidium bromide, acridine orange, and sodium dodecyl sulfate) and by the high rate of spontaneous loss of r-determinants. Dyebuoyant density centrifugation allowed recovery of R-plasmid DNA that measured roughly 24 mum in contour length; after 'curing' with concomitant loss of 9 r-determinants, the contour length of the R-plasmid DNA of one isolate (No. SE 154) had decreased to roughly 15 mum, and none was detected in the sole variant of the isolate that spontaneously had lost 11 r-determinants.


A clinical investigation of pivmecillinam. A novel beta-lactam antibiotic in the treatment of urinary tract infections. Pivmecillinam (FL 1039) is the pivaloyloxymethyl ester of mecillinam (FL 1060) which has considerable in vitro activity against Enterobacteriaceae. 38 hospital inpatients who had proven urinary tract infections were treated with 400 mg pivmecillinam four times daily for 5-7 days. The MIC of mecillinam to the infecting organisms was determined as were the serum and urinary concentrations of the antibiotic. The patients were followed up for 4-6 weeks after the end of treatment. Three patients were lost to follow-up. Of the 35 patients who were adequately followed up, 29 (83%) were classified as cured and there were 6 failures. Reported side effects were of a minor nature.


Infection of newborn Taiwan monkeys (Macaca cyclopis) with human adenovirus type 12 and simian virus 40. For testing biological response of newborn Taiwan monkeys to infection of human adenovirus type 12 (Ad12) and simian virus 40 (SV40), 40 newborn Taiwan monkeys were inoculated with Ad12, Ad12, plus SV40, Ad12 supplemented with Ad12-induced hamster tumor tissue (Ad12 tumor) or control specimens (HeLa or African green monkey kidney cell lysate). Among them 26 survived including 8 newborn monkeys inoculated with control specimens. The survivors were observed for 4 years but no tumor was produced. The increase of body weight and intake of calories and protein in each test group during the first 12 wk were similar to those of corresponding control groups. Intrapulmonary inoculation of 108.2TCD50 of Ad12 with additional subcutaneous dose of Ad12 (108.8TCD50), Ad12 plus SV40 (108TCD50) or Ad12 plus Ad12 tumor killed 78% of newborn monkeys (7 of 9) in 18 days. The newborn could stand subcutaneous inoculation of SV40 (108TCD50) with 1 dose of Ad12 (108.8TCD50) or 3 doses of Ad12 (108.5, 108.5 and 108.2TCD50) at 24-hr intervals. When 108.8TCD50 or more Ad12 were inoculated, the virus could be isolated as late as 44 and 26 days from rectal and throat swab specimens respectively. The Ad12 neutralizing antibody in baby monkeys inoculated with multiple doses of Ad12 persisted, in low titer, longer than those injected with single high doses of Ad12, but anti-Ad12 T (tumor) antibody disappeared by 35 wk in both groups. Although SV40 antibody response was better than Ad12 antibody response in baby Taiwan monkeys, pre-infection of SV40 did not potentiate the production of anti-Ad12 T antibody.


Host preference of mosquito vectors of Japanese encephalitis. The host preference of 4 Culex mosquito species collected in Miaoli and Pingtung counties, Taiwan was studied by capillary precipitin method. Antisera to alum-precipitated sera of man, bovine, swine, rabbit, horse, dog, cat, mouse, chicken, duck, and pigeon were produced in rabbits and reacted with 758 mosquito blood meals among which reactions to one or more antisera. Culex annulus and Culex tritaeniorhynchus summorosus showed a great avidity for pig, and Culex fuscocephala for bovine. Culex pipiens fatigans was ornithophilic. None of 110 C. t. summorosus and 2.4% of 223 C. annulus had fed on man. Among 66 samples of C.p. fatigans tested 10.3% had fed on man, while none of 359 C. fuscocephala did. It seems that the latter does not act as a primary vector of Japanese encephalitis.


Permeability of pulmonary vascular endothelium. Three aspects of transendothelial exchange in the lungs are considered: stretching of interendothelial junctions of pulmonary microvessels by increase in pulmonary capillary pressures; selective stretching of interendothelial junctions of bronchial venules in response to histamine, bradykinin and endotoxin; active transport of peptides across the body of the endothelial cell after enzymic action at or near the luminal surface of the endothelial cell. Stretching of interendothelial junctions between the cells lining the pulmonary capillaries was demonstrated using a variety of macromolecular tracers under controlled haemodynamic conditions. Selective leakage of bronchial venules, the systemic venules of the lungs, was shown using colloidal carbon as a tracer. Transendothelial transport of peptides across the pulmonary capillary lining involved the use of electron microscopic autoradiography after intravenous administration of radioactively-labelled lipoproteins. Different mechanisms appear to provide routes of entry into the perivascular interstitial spaces of the lungs.