Nasopharyngeal carcinoma and Burkitt's lymphoma in a Canadian family. I. HLA typing, EBV antibodies and serum immunoglobulins. Two nasopharyngeal carcinomas of the lymphoepithelioma type and two Burkitt's lymphomas with the characteristic histopathologic features developed in three siblings and one first-degree cousin in a large French-Canadian family. Epstein-Barr virus antibody titres in the two lymphoepithelioma cases but not in the Burkitt's lymphoma cases were, as expected, greatly elevated. HLA typing of the family members failed to disclose HLA antigens A2 and B Sin-2, which have been associated with lymphoepithelioma in Asia. The occurrence, however, of a plasmacytoma in one other first-degree cousin and low serum IgA values in several siblings and cousins suggests the possibility of a genetically determined predisposing B-cell dysfunction in the development of these tumours.


Nephrotoxic and cytoproliferative effects of streptozotocin: report of a patient with multiple hormone-secreting islet cell carcinoma. The nephrotoxic and cytoproliferative side effects observed in a patient with Streptozotocin-treated, multiple hormone-secreting, pancreatic islet cell carcinoma are described. Streptozotocin induced prolonged partial remission of the patient's multiendocrine syndrome but resulted in progressive azotemia, which was controlled by temporary hemodialysis. A renal biopsy, the first to be reported in detail in such a condition, demonstrated a tubulo-interstitial nephritis and a glomerular alteration consisting of cellular nodules. At autopsy there were numerous bilateral renal cortical spindle cell "tumors" and cellular aggregates in glomeruli. These findings suggest that the tumorigenic effects of Streptozotocin demonstrated in animals may also occur in man.


Effect of oral prophylactic broad spectrum nonabsorbable antibiotics on the gastrointestinal absorption of nutrients and methotrexate in small cell bronchogenic carcinoma patients. Patients with small cell bronchogenic carcinoma, in a study utilizing laminar-air-flow-protected environments, oral prophylactic broad spectrum non-absorbable antibiotics (PNAA), and intensive combination chemotherapy, were examined to determine the effects of PNAA on serum biochemical values and on gastrointestinal absorption of both nutrients and methotrexate. With use of PNAA the following abnormalities were observed; serum carotene and folate decreased, D-xylose absorption was impaired, fat globules and muscle fibers were demonstrable in the stool, and the mean weight loss in 6 weeks was 10.2% as compared with 4.3% in patients not treated with antibiotics. Methotrexate absorption decreased from a mean of 69% prior to antibiotic use to 44% on PNAA. Thus, PNAA causes malabsorption of both nutrients and drugs. It appears unwise to treat patients on PNAA with oral antineoplastic drugs. Nutritional status must also be closely monitored and supplemental nutrition, either intravenously or with elemental diets, must be considered.


Small cell nonkeratinizing carcinoma of the cervix associated with ACTH production. The case of a 40-year-old woman with primary small cell carcinoma of the cervix is reported. She developed widespread metastates and florid Cushing's syndrome. Serum ACTH levels were greatly elevated and no site of production other than the tumor could be demonstrated at autopsy. The tumor cells demonstrated features characteristic of cells of the APUD series. Such cells have been demonstrated in normal cervical epithelium; it is likely that they may become malignant, giving rise to tumors with a potential to secrete polypeptide hormones. This case suggests that endocrine active "Apudomas" may arise from the uterine cervix. Certainly, patients with small cell carcinoma of the cervix should be investigated with appropriate serum assays for polypeptide hormones.


Ultrastructural, histochemical, and biochemical studies of two cases with amylase, ACTH, and beta-MSH producing tumor. Two cases of intrathoracic tumor, different in histology and accompanied by hyperamylasemia, were studied ultrastructurally, histochemically, and biochemically. The ultrastructure of the tumor cell cytoplasm showed many zymogen granules in case 1 and smaller cored granules in addition to zymogen granules in case 2. Both tumors contained not only a large amount of amylase, which was electrophoretically of saliva type with three components, but also significant amounts of immunoreactive ACTH and beta-MSH. Starch film and immunofluorescence showed that the tumor cells stored amylase. It was concluded from these findings that the tumor cells ectopically producing amylase, which showed differentiation toward the cells with zymogen production, could differentiate toward the cells of ACTH-MSH system at the same time.


Carcinosarcoma and mixed müllerian tumors of the fallopian tube: report of four cases. We report a 74-year-old woman with malignant mixed müllerian tumor and three women, aged 47, 58, and 76 years, with carcinosarcomas, all primary in the fallopian tube. The tumors grew predominantly intraluminally and were associated with hydrosalpinx. All four patients underwent salpingectomy. Two of them received radiation therapy; one died within 9 months because of the tumor and the second is well 2 years after diagnosis. Of the two women without additional therapy, one is alive with tumor 53 months postoperatively and the other is well 1 year after surgery. Available data suggest that these neoplasms are relatively radioresistant and that their prognosis correlates best with local invasiveness.


Carcinoma of the stomach in childhood.

Carcinoma of the stomach in childhood. A case of mucinous adenocarcinoma of the stomach is described in a 20-month-old child with a large abdominal mass. The lesion arose from the lesser curvature of the stomach and invaded the submucosa and lymphatic channels. A partial response to chemotherapy, including vincristine, Cytoxan, actinomycin D, and adriamycin, was noted, but the patient died 3 1/2 months following diagnosis shortly after gastric perforation and peritonitis developed. Autopsy revealed a pattern of widespread visceral metastases, accompanied by pathologic findings suggestive of ataxia-telangiectasia. A diagnosis of mucinous adenocarcinoma, although it is rare, should be considered in the evaluation of primary neoplasms of the stomach in children.


The morphology of tumors of the human gastrointestinal tract in short-term organ culture and the reaction of these tumors to infection with poliovirus. A morphologic study of 57 cancers of the human gastrointestinal tract maintained in short-term organ culture was carried out, and the response of part of these tumors to infection with poliovirus was studied. The tumors, grown in vitro, mainly retained their original characteristics and histologic types. A certain correlation between the histologic type and its viability in short-term organ culture was revealed: the more differentiated forms were maintained better than the poorly differentiated ones. A comparative study of the polio-virus-infected and the control cultures indicated that wild poliovirus showed the most marked oncolytic activity in short-term organ culture, whereas attenuated poliovirus III produced a less marked, although still distinct, destructive effect.


Lymph node involvement by direct extension in adenoid cystic carcinoma. Absence of classic embolic lymph node metastasis. Thirty-four cases of adenoid cyctic carcinoma seen at the University of Virginia Hospital from 1946 to 1974 were reviewed, with special emphasis on lymph node involvement by tumor. Lymph node involvement was found in three cases of primary tumors of the submaxillary gland, and all of the affected lymph nodes were in the immediate vicinity of the primary tumor. Two lymph nodes were involved in two of the cases, and one node was involved in the third case. In all of these lymph nodes, adenoid cystic carcinoma was present in the soft tissue surrounding the node, and the tumor extended into the node. No metastatic tumors were observed in 46 lymph nodes removed incidentally at the time of local excision of the primary tumors in 10 additional cases or in 212 lymph nodes examined after unilateral radical neck dissections in six other cases. Five autopsies in this series showed no lymph node metastases. In this series of cases adenoid cystic carcinoma only invades lymph nodes in the immediate vicinity of the primary tumor. When lymph node involvement does occur, it does not result from embolic lymph node metastasis; rather, a direct invasion of the lymph node from tumor in the perinodal soft tissue occurs. Obviously, this small study does not completely exclude the possibility of embolic metastasis; however, if it does occur, it must be extremely rare.


Hodgkin's disease and granulomatous angiitis of the central nervous system. Granulomatous angiitis of the central nervous system, a pathologic entity associated previously with a fatal prognosis, is reported in a patient with Hodgkin's disease. Viral isolation, indirect fluorescent antibody, and electron microscopic studies performed on fresh cerebral tissue were negative. The granulomatous angiitis of the central nervous system in this patient remitted following therapy for Hodgkin's disease. The epidemiologic, clinical and pathologic data in all reported cases of granulomatous angiitis of the central nervous system suggest that it: 1) may have more than one etiology; 2) may not always be fatal; 3) is associated with varicella-zoster virus and Hodgkin's disease; 4) is one of two granulomatous reactions found in association with Hodgkin's disease; and when found with Hodgkin's disease, 5) may remit with adequate therapy for the lymphoma.


An immunoelectron microscopic analysis of Epstein-Barr virus-associated complement-fixing antigen. Epstein-Barr virus-associated, complement-fixing antigens have been observed with a complement-mediating immunoreaction and with the peroxidase-labeled anticomplement antibody electron microscopic method. Postive reaction products could be found in the nuclei of P3HR-1 cell lines. At high magnification, it was ascertained that the reaction-positive precipitates were associated with chromatin and were mostly either finely granular or filamentous, which suggests that the antigen was not uniform. It was speculated that the antigen would be analogous to the SV40 T-antigen, since the localization pattern of the positive reaction was similar in both antigens. This new, modified method using complement may also be used for detection of a natural antibody of unknown class or for low-sensitivity systems of antigen-antibody reactions. Thus, this method appears useful for studying various kinds of experimental materials.


Radical partial renal irradiation. an alternative to partial nephrectomy in bilateral Wilms' tumor. A 6-year-old girl with simultaneous bilateral Wilms' tumor was treated with a radical nephrectomy on the side of more extensive involvement, followed by postoperative renal bed irradiation (3500 rads over 32 days) and radical partial renal irradiation (4000 rads over 35 days) to the contralateral disease. Concurrent and subsequent adjuvant actinomycin-D and vincristine were given. She has remained disease-free for over 5 years after primary therapy, with satisfactory renal function. This case report suggest that radical partial renal irradiation may be an alternative to partial nephrectomy in the management of the lesser involved kidney in some patients with bilateral Wilms' tumor.


Calcium metabolism in cancer. Studies using calcium isotopes and immunoassays for parathyroid hormone and calcitonin. Studies of calcium metabolism in 38 patients with cancer indicated that: 1) intestinal absorption of calcium was reduced in patients with skeletal metastases and in those with hypercalcemia; 2) calcium-47 space (a measurement of bone turnover rate) was high in the patients with skeletal metastases; 3) hypercalcemic patients had higher urinary and endogenous fecal excretion of calcium than those who were normocalcemic; 4) levels of plasma immunoreactive parathyroid hormone were similar in normo- and hypercalcemic patients, but the levels for a given serum calcium in malignant disease were lower than those in primary hyperparathyroidism; and 5) some patients had elevated calcitonin levels. Hypercalcemia complicating malignant disease is therefore not due to hyperabsorption or diminished excretion of calcium, and a low calcium diet is unlikely to benefit these patients. Measurement of 47Ca space could be of use in monitoring therapy of patients with skeletal metastases, and measurement of plasma parathyroid hormone could be useful in the differential diagnosis of hypercalcemia.


Cyclic AMP as a determinant of vulnerability to ventricular fibrillation in the isolated rat heart. Ventricular fibrillation threshold and vulnerable period were measured in the isolated perfused rat heart to assess the influence of dibutyryl cyclic AMP on ventricular vulnerability. Doses of dibutyryl cyclic AMP of 0.6-1.6  mumol/min caused an increase in coronary flow but had no effect on vulnerability, whereas doses of 2.6-4.2 mumol/min resulted in an increase in coronary flow, a decrease in VF threshold, and an increase in the width of the vulnerable period. These experiments support the concept of a local myocardial action of catecholamines, mediated by cyclic AMP, whereby vulnerability to ventricular fibrillation is increased.


[Polymorphism of granular endoplasmic reticulum in the ovocytes of Mytilus edulis L]. In the cytoplasm of the ovocytes of Mytilus edulis, at the beginning of the yolk elaboration, but especially at the end of vitellogenesis, the polymorphous rough endoplasmic reticulum is very well developed. The concentrical disposition of the cisternae around of the mitochondria, the lipid inclusions and some granules of the yolk, are very frequent and must be considered as a system for increasing the area of proteic synthesis.


[Spontaneous and induced sterility by ethyl methanesulfonate (EMS) in Nigella damascena L]. EMS-induced sterility could be very partially due to chromosomal aberrations appearing during male and female meiosis or even to mechanical abnormalities of the double fertilization. The sterility could be also related to diplontic origin (lethal factors or small deficiencies appearing in homozygous state on account of self-pollinization). Owing to our histological and genetical data, a female gametophytic origin could be mainly ascribed to EMS induced sterility. It could arise from a damage of the feeding function of the nucellus.


Alterations in the neurohypophysis of rats treated with chlorphentermine or tricyclic antidepressants. Rats were treated with several amphiphilic, cationic compounds that are known to cause generalized lipidosis (chlorphentermine, iprindole, 1-chloro-amitriptyline, clomipramine). After prolonged drug treatment the neurohypophysis showed severe morphologic alterations particularly in Herring bodies (HB), perivascular cells, and pituicytes. HBs displayed the following abnormalities: (a) great accumulation of autophagic vacuoles that contained neurosecretory granules (NSG); (b) numerous coarse osmiophilic conglomerates; (c) masses of multilamellated material; (d) reduced numbers of intact NSGs. Perivascular cells accumulated large lamellated inclusion bodies. Pituicytes contained membrane-bound crystalloid inclusion bodies. The noxious effect of chlorphentermine and l-chloro-amitriptyline was more pronounced than that of iprindole and clomipramine. The alterations in perivascular cells and in pituicytes are typical of drug-induced lipidosis. The lesions in HBs are tentatively explained as follows: HBs were previously proposed to be the sites of normally occurring intraaxonal disposal of excess neurosecretory material. The present experimental conditions interfere with this catabolic process. Incomplete digestion of the axo-plasmic constituents due for disposal might result in abnormal accumulation of NSG-containing autophagic vacuoles, osmiophilic conglomerates, and multilamellated material. This eventually leads to degeneration of HBs. The functional implications of the neurohypophysial lesions remain to be elucidated by functional experiments.


Effect of collagenase on mechanical activity and fine structure of an intestinal smooth muscle. The effect of the enzyme collagenase (40-200 units-ml-1) on the spontaneous mechanical activity in vitro and on the fine structure of the activity of the taenia was enhanced both in the isometric and isotonic recordings; after several minutes the muscles became slack or elongated to up to twice their resting lengths. The structural changes were dramatic but a number of muscle cells remained apparently unaltered even with the highest concentration and the longest incubation time (120 minutes). The large variety of structural changes were tentatively grouped into two separate sequences. One sequence involved swelling of the muscle cell, dispersion of the filaments and breaking up of the cell membrane: the thick myofilaments increased considerably in size and became heterogeneous in size and shape, but were still recognizable after disruption of the cell membrane. The other disruptive sequence involved separation of the superficial part of the muscle cell, which became electron-lucent, from the core of the cell where filaments were very densely packed. Few or no changes were observed in non-muscle cells.


Extraneuronal effects of 6-hydroxydopamine and extraneuronal uptake of noradrenaline. In-vivo and in-vitro studies on adrenocortical cells of lizards and rats. 6-hydroxydopamine (6-OHDA) was shown to cause ultrastructural changes in adrenocortical cells of lizards and rats. These changes comprised the formation of dense bodies with lamellar and crystalloid patterns, a decrease in the number of mitochondria and structural alterations of mitochondria. Alterations in adrenomedullary cells were not affected as a rule. Only in young animals did 6-OHDA cause deposits of an electron-dense material in medullary cells. An attempt was made to obtain information on amine uptake into cortical cells using the Falck-Hillarp technique to analyse the in-vivo and in-vitro uptake of noradrenaline (NA) into the adrenal cortex in adult rats. Extraneuronal uptake into heart and spleen was studied as well. Our results suggest that NA is taken up into cortical cells, particularly into nuclei, after exposure to 10(-4) gm/ml in-vitro indicating that uptake of 6-OHDA is also likely. Investigations using labelled 6-OHDA is also likely. Investigations using labelled 6-OHDA are required for further elucidating its extraneuronal uptake.


A multilamellate body in the granulated hypophysial cells of the teleost, Hemihaplochromis philander. A multilamellate body (MLB), bearing close resemblance to an array of annulate lamellae, has been observed in several adenohypophysial cell types of the teleost, Hemihaplochromis philander. In longitudinal section, each MLB comprises a ladder-like row of 12-50 sausage-shaped profiles, termed lamellae. A few lamellae in each section show connections with the endoplasmic reticulum. Apposition of paired lamellar membranes at regularly spaced intervals results in a beaded appearance, whereby sites of membrane apposition are probable pore sites. The MLB differs from annulate lamellae in having poorly developed pores and closer packing of lamellae. It is suggested that the MLBs described here, may represent annulate lamellae at a stage of development or break-down when pores are incomplete.


Exocrine pancreas under experimental conditions. II. In vitro formation of paracrystalline structures and its functional consequences. The in vitro formation of paracrystalline structures after addition of high concentrations of amino acids to the incubation medium was investigated in pancreatic lobules and pancreatic homogenates. It could be shown that even in the homogenate, pCPA induced the formation of paracrystalline structures which exhibited the same ultrastructural arrangement as seen in the in vivo induced paracrystalline inclusions in the RER of the rat pancreas. In correlation with the morphological alterations, the functional consequences to the secretory process, i.e. amylase discharge,discharge of pulse-labeled proteins and incorporation of 3H-leucine into pancreatic proteins,were studied in pancreaticlobules. Two different approaches were used. Firstly, the effect of pCPA-pretreatment of rats and secondly, the effect of higher pCPA concentrations in the incubation medium on the secretory process in untreated pancreatic lobules were studied. A nonparallelism of inhibition of the three different steps of the secretory process depending, with respect to its extent,on time after pCPA-application (4-72 h) and on the concentration of pCPA (1-10(-5) to 1-10(-2) M) in the incubation medium was found. In addition to specific effects probably due to pCPA and to the paracrystalline inclusions, unspecific alterations, particularly accompanying degenerative processes after in vivo pretreatment, could be differentiated.


Extraneuronal effects of 6-hydroxydopamine. Tissue culture studies on adrenocortical cells of rats. The effects of various concentrations of 6-hydroxydopamine (6-OHDA) on rat adrenocortical cells in tissue culture were studied with phase contrast and electron microscopy. With 40 mg/l of 6-OHDA the first signs of alteration as revealed by microcinematography appeared in isolated cortical cells as early as 15 min after addition of the drug. There was a cessation of movement of cell organelles and an immobilisation of membrane undulations followed by the development of dark inclusion bodies. The cells underwent increasing shrinkage and collapsed by 1 1/2 h. Chromaffin cells were not affected until 45 min after exposure to the drug and neurons were the most resistant population. However 6 1/2 h after application of the drug most cells in the culture were dead. 6-OHDA applied in different doses and to adrenal explants did not alter the sequence of events. Ultrastructurally cortex cells underwent damage alone two lines: they either showed lytic changes or developed various types of dense bodies before reaching the lytic stage. Treatment of cortical cells with 40 mg/l 5-or 6-OHDA followed by exposure to buffered 2% glyoxylic acid and heat did not produce a fluorescence within the cells. Microspectrofluorimetry on amine models with noradrenaline, 5- and 6-OHDA revealed that neither 5-nor 6-OHDA are capable to form a fluorophore with glyoxylic acid.


Location of the sequences coding for capsid proteins VP1 and VP2 on polyoma virus DNA. The 19S and 16S polyoma virus late mRNAs have been separated on sucrose-formamide density gradients and translated in vitro. The 16S RNA codes only for polyoma capsid protein VP1, while the 19S RNA codes in addition for capsid protein VP2. Since the 19S and 16S species have been previously mapped on the viral genome, these results allow us to deduce the location of the sequences coding for VP1 and VP2. Comparison of the chain lengths of the capsid proteins with the size of the viral mRNAs coding for them suggests that VP1 and VP2 are entirely virus-coded. Purified polyoma 19S RNA directs the synthesis of very little VP1 in vitro, although it contains all the sequences required to code for the protein. The initiation site for VP1 synthesis which is located at an internal position on the messenger is probably inactive either because it is inaccessible or because it lacks an adjacent "capped" 5' terminus. Similar inactive internal initiation sites have been reported for other eucarotic viral mRNAs (for example, Semliki forest virus, Brome mosaic virus, and tobacco mosaic virus), suggesting that while eucaryotic mRNAs may have more than one initiation site for protein synthesis, only those sites nearer the 5' terminus of the mRNA are active.


Homologous synthesis: a method for the preparation of mixed substrates and individual compounds. Equimolar mixtures of several homologous or vinylogous alkyl methanesulfonates (mesylates) are converted, by a series of reactions, into equimolar mixtures of compounds having one methylene group more, per molecule, than the starting material. Such mixtures can be used for studying the specificity of an enzymatic reaction in a single experiment in vivo or in vitro. The simultaneous synthesis of a mixture of compounds is also of great advantage in the preparation of a series of radioactively labelled substances. It is more convenient and much more efficient to prepare several labelled compounds of a homologous or vinylogous series in a single reaction, and to isolate each of them by chromatographic methods, than to synthesize the individual substances separately.


Identification of isomeric doxyl stearic acids by gas-liquid chromatography and mass spectrometry. Direct probe and GC/MS spectra were determined for the isomeric 4- to 16-doxyl stearic acids and their methyl and silyl esters in pure form and in mixture with natural fatty acids and their esters. The base peak for all free and esterified doxyl stearic acids was at m/e 281. The methyl esters of all isomers gave nearly identical fragments in the high mass regions having M+ at m/e 398 with intensities of 2-3%. The isomers were identified on the basis of the fragments retaining the doxyl group, which had positive charge and were different for each compound. It was shown that the fragment m/e 281 may be used to identify and quantitate the stearate derivatives in presence of natural fatty acids. The silyl esters of the doxyl stearates gave complex mass spectra. The isomeric doxyl stearates were resolved by GLC on 3 ft. glass columns containing 1% SE-30 packing as methyl esters.


Isolation of a novel lipid hapten, cytolipin S, from rat spleen. Antisera against rat erythrocytes contain agglutinins directed against unknown lipid determinants. Complement-fixation shows more reactivity with lipid extracts of rat spleen than of other rat tissues. The isolation of the reactive lipid from rat spleen, cytolipin S, is described. Cytolipin S is a glycosphingolipid containing glucose, galactose, and galactosamine with molar ratios of 1:2:1. It migrates on TLC like asialo GM1 (more slowly than cytolipin R, a ceramide tetrasaccharide, or cytolipin F, a ceramide pentasaccharide). Asialo GM1 and cytolipin S, when properly combined with auxiliary lipids, react very similarly with anti-rat erythrocyte sera by complement-fixation. However, cytolipin S is much more effective than asialo GM1 in inhibiting the hemagglutination reactions. It is concluded that cytolipin S and asialo GM1 are ceramide tetrasaccharides having different chemical structures and that the structural differences probably will be found in the carbohydrate linkages other than that between the terminal and penultimate residues.


An ESR study of the hydration steps of lecithin multilayers. A spin-labeled stearic acid and other spin-labeled molecules have been used for an ESR study of the hydration process of the egg lecithin lamellar phase for water concentrations, CW greater than 10%. Two hydration steps are found for CW approximately 20% and CW congruent to 30%, in good agreement with results of the literature. To explain discontinuities in the behavior of three spin-labeled molecules at CW congruent to 20%, a conformational change of the phosphatidylcholine polar head and a phase transition in the polar interface are proposed.


Potency of field samples of oral poliovirus vaccine. The transport and storage facilities and the potency of oral poliovirus vaccines currently administered in 108 centres in India were investigated. Storage and distribution practices in many of the centres were far from ideal. There was no significant loss of potency in the vaccine samples collected from a few centres, while samples from other centres showed a 60-99% loss of virus particles per dose. A national monitoring system has since been established to check the potency of every batch of oral poliovirus vaccine imported and to streamline the transport, storage, and administration of the vaccine. Constant vigilance as regards the quality of vaccines should ensure the success of any poliomyelitis immunization campaign.


Some characteristics of poliovirus strains isolated in Uganda between 1966 and 1971. Sixty-five poliovirus strains were investigated in genetic marker tests in order to obtain information on the characteristics of polioviruses circulating in Uganda where, owing to the insufficient use of live poliovirus vaccine, poliomyelitis remained a serious public health problem. Of the type 1 strains predominant in both epidemic and non-epidemic years, 29 were studied for their antigenic fine structure. Based on their intratypic character, these strains proved to represent six different antigenic variants. Three of these variants were predominant during certain periods; the first variant was present in 1966 and 1968, the second in 1967, and the third from 1969 to the end of observation period. Four strains from Kuwait and three from Ghana isolated in 1969 and 1970 showed an antigenic structure identical to that of the strains predominant in Uganda in these years. Some strains proved to be of vaccine origin. Twenty-nine type 1 and 24 type 2 strains showed a great variety of characteristics when studied in d, od, and rct/40 marker tests. There was no indication that the distribution of strains according to their in vitro markers would have been different in epidemic and non-epidemic years, or that any particular combination of markers would have been more common among strains isolated from paralytic patients than among those from non-paralytic patients. Nine of 12 type 3 strains had the rct/40(+) marker.


Monkeypox and whitepox viruses in West and Central Africa. Prospects for the eradication of smallpox are now highly encouraging. With the cessation of man-to-man transmission, the question of possible animal reservoirs of smallpox becomes increasingly important. During the period 1970-1975, 20 cases of a smallpox-like disease were detected in smallpox-free areas of tropical rain forest in West and Central Africa. Epidemiological and virological investigations revealed that the disease was caused by an animal poxvirus termed monkeypox virus, a member of the orthopox virus group. The disease spread with difficulty even among susceptible close contacts and does not appear to be sufficiently transmissible to permit continuing infection to become established in man. During the investigations, four orthopox viruses termed whitepox viruses were isolated from rodents and monkeys. The isolates were not distinguishable from variola virus with currently available laboratory techniques, but there is no evidence so far that viruses of this group have infected man. Although there is now substantial and accumulating evidence that there is no animal reservoir for smallpox, continued surveillance and studies in West and Central Africa are warranted.


[Anatomo-pathologic aspects of phycomycoses. 6 clinical cases observed in Ivory Coast]. Six cases of superficial phycomycosis have been recently demonstrated in Ivory Coast, the total number of cases being therefore 8 in this country. After a brief review of the etiology, clinical studies, treatment, and geographical distribution of the 150 world cases known to-day, the author studies the histological lesions of this hypodermic granulatous inflammation, whose characteristic aspects (polynuclear micro-abcesses, palissade histiocytes, filaments sheathed in an eosinophil precipitate) are easy to recognise. These anatomopathological aspects vary according to the topography, clinical evolution and treatment. The author presents a summary of the only ultrastructure known today. He points out the significance of this anatomo-pathologic study, which shows the evolution of the immunitary reaction and permits a relatively precise pronostic.


Monkeypox-specific antibodies in human and simian sera from the Ivory Coast and Nigeria. A test for monkeypox-specific antibodies is described. Monkeypox immune sera can be made type-specific by immunoabsorption with heterotypic poxvirus extracts. Monkeypox-specific antibodies were demonstrated in sera from 9 cynomolgus monkeys (Macaca fascicularis) that had previously been experimentally infected with monkeypox. Monkeypox-specific antibodies were found in 3 wild-caught African monkeys (Cercopithecus spp.) and in 3 human sera collected from Africans in the Ivory Coast and Nigeria 3(1/2)-4 years after they had suffered a pox-like infection. Monkeypox had been recognized in one of the patients by virus isolation, and had been suspected in the others for epidemiological reasons. Vaccinia-specific antibodies were found in 4 human sera collected 6 weeks after smallpox vaccination.The serological results provide the first laboratory evidence of a monkeypox reservoir in wild monkeys.


[Control of the regeneration of small pieces of the planarian Dugesia tigrina by cylic AMP and GMP nucleotides]. The effects on regeneration of cyclic nucleotides AMP and GMP were studied on small pieces of the planarian worm Dugesia tigrina. db cAMP, 8Br cGMP and substances acting on their metabolism were used in this work (aminophylline, insulin, imidazole). It seems that these second messengers play a role in a mechanism of regulation in which cGMP has a promotive action and cAMP an inhibitive one.


[Electron microscopic observation of the infectious cycle of Visna virus in sheep choroid plexus cells at temperatures under 37 degrees C]. Electron microscope study of viral penetration at 4 degrees C shows that viral particles first attach to cells. The attachment sites are invaginations which later fuse to form phagocytic cytoplasmic vacuoles. Some virus particles with altered envelopes can be seen in the cytoplasm. After ten days of incubation at 29 degrees C, filamentous inclusions and mature viral particles are seen in the cytoplasm. The cellular membrane budding process and virus release observed at 37 degrees C have disappeared. When the above preparations are warmed to 37 degrees C, budding is seen, and two kinds of extracellular particles are present, one with a clear center and the other with a dense core (mature virus).


[Immunocytochemical evidence for the binding of insulin to the somatotrophic cells of the human hypophysis]. An immunocytochemical method has demonstrated a hypophyseal cell surface fixation of molecules exhibiting immunological properties of insulin. This fixation occurs massively on GH-cells and on a minority of ACTH cells of the human antehypophysis.


[Statistical relationship between "slow-wave sleep" and "paradoxical sleep" in rats]. In the Rat, statistical analysis of slow sleep (SS) and paradoxical sleep (PS) episodes disclosed two significant (p less than 0,01) positive correlations: (1) Over a four-hour period, the mean duration of PS episodes was correlated to the mean of the just preceding "light" SS episodes; (2) On the contrary, from cycle to cycle of sleep-wakefulness, the duration of each PS episode was correlated to that of the "light" SS episode of the next cycle.


[Cytoimmunological study of the corticotrophic and corticomelanotrophic cells in the adenohypophysis of Boops salpa L. (marine teleost) in different experimental conditions (variation of salinity and of background color)]. The cytoimmunological study of the adenohypophysis of Boops salpa (marine Teleost), in different experimental conditions confirmed that the MacConaill's hematoxyline-positive cells contain: 1. ACTH (Corticotrophic Hormone) in the pars distalis, 2. MSH (Melanocyte-Stimulating Hormone) and ACTH in the pars intermedia.


[Pharmacologic properties of the toxins isolated from the sea anemone (Anemonia sulcata)]. We have shown that if toxin II isolated from Anemonia sulcata is mainly cardiotoxic on Rats it nevertheless displays neurotoxic effects. Furthermore it releases acetylcholine from isolated Guinea Pig ileum.


Detection of perioperative myocardial damage after coronary artery bypass graft surgery. In order to evaluate methods for detecting peri-operative myocardial damage we studied 41 patients before and serially following coronary artery bypass graft surgery utilizing the 12-lead ECG, serum MB-CPK measurements, and 99mTc pyrophosphate myocardial scans. Six of the 41 patients (15%) developed persistent new Q waves after surgery. Six other patients demonstrated ischemic ST-T wave changes that persisted for 48 hours or more. Mean total MB-CPK released was highest for the group with new Q waves [1598+/-545 (SE) I.U./L-hr] as compared to the group with ischemic ST-T wave changes 708+/-65 I.U./L-hr) or the group with no ECG changes (262+/-47 I.U./L-hr). Ten patients (24%) has positive postoperative pyrophosphate scans consistent with myocardial infarction. The three techniques were compared in these 41 patients utilizing 465 I.U./L.-hr as the upper limit of normal MB-CPK released after uncomplicated coronary bypass surgery (no ECG changes, negative scan). Five patients with ischemic ECG changes had a positive scan and high MB-CPK; six patients with no ECG changes had high MB-CPK but a negative scan; and one patient with high MB-CPK and new Q wave had a negative scan. We conclude 1) new Q waves on ECG underestimate the incidence of myocardial damage after coronary artery surgery; 2) MB-CPK alone overestimates the incidence of infarction; and 3) a combination of the three techniques is the best means for detecting myocardial damage after coronary artery bypass graft surgery.


The characterization of the ACTH produced by a primary pituitary tumour in a patient with Cushing's disease. The nature of ACTH present in pituitary tumours associated with Cushing's disease has not been previously characterized nor correlated with the electron microscopic appearance. The present report describes the culture of tumour tissue obtained from a patient with a pituitary tumour associated with Cushing's disease, and the characterization of the ACTH content of tumour and media by bioassay, immunoassay, and Sephadex G-50 gel filtration. Electron microscopic studies were also performed. The pathological diagnosis was pituitary adenoma with basophilic PAS-positive granules. Electron microscopy showed uniformity of size and shape of the tumour cells, the presence of secretory granules of varying size and density, and disorganized tubular and vacuolar arrays of endoplasmic reticulum. The bioreactive ACTH content of the tumour was 0-29 mug/g, which is markedly below that seen in the normal pituitary, but within the range reported for ectopic ACTH-producing tumours. Immunoreactive ACTH when measured by a C-terminal antibody was five-fold higher than when measured by an N-terminal antibody; the latter gave a value of 1-24 mjg/g. On Sephadex G-50 gel filtration, 9% of the N-terminal immunoreactivity was present in the Vo fraction ('big' ACTH). This latter fraction had a greater percentage of bioreactivity (28%) than previously reported for this molecular species. Analysis of the tumour culture medium revealed a variation in molecular size similar to that seen in the tumour, although the percentage of ACTH of large molecular size was greater, suggesting increased secretion of a possible 'prohormone' by the tumour. Plasma ACTH was characterized by a 2:1 ratio of immunoreactivity (N-terminal) to bioreactivity, and a 4:1 ratio of C-terminal/N-terminal immunoreactivity. This report also appears to be the first of successful short-term tissue culture of a primary ACTH-producing tumour. The granule size was considerably larger than that reported for normal pituitary ACTH-containing cells.


Hormonal variations in secondary amenorrhoea during the metyrapone test. The occurrence of uterine bleeding following administration of Metyrapone in cases of secondary amenorrhoea and oligomenorrhoea led the authors to measure pituitary, gonadal and adrenal hormone levels, before and after the oral administration of this drug in sixteen patients, in an attempt to provide an interpretation of this phenomenon. There were expected changes in plasma ACTH and urinary 17-hydroxycorticosteroids, but no variations were noted in serum FSH and LH, plasma oestradiol and urinary oestrogens; the most important alterations occurred in plasma progesterone and urinary pregnanetriol levels. It is suggested that bleeding occurs because of the changes which are induced in plasma progesterone levels.


1-Alpha-hydroxycholecalciferol in the treatment of hypocalcaemic psychosis. Two patients with profound hypocalcaemia due to uraemia and hypoparathyroidism respectively presented with alterations of mental state as their dominant symptom. 1-Alpha-hydroxycholecalciferol (1-alpha-OHD3) was chosen as the principal therapy because of its potency and rapid action. In both patients the plasma calcium was restored towards normal and the symptoms relieved within a very short period. The cases illustrate the role of 1-alpha-OHD3 in severe hypocalcaemia due to two different causes and provide experience on which to base treatment regimes for future patients similarly afflicted.


The effect of electric convulsion therapy on the circulating concentrations of pituitary hormones, cortisol and cyclic adenosine monophosphate. In order to evaluate the effect of electroconvulsive therapy on the human pituitary-adrenal function, serial plasma samples were assayed for FSH, LH, TSH, HGH, ACTH, cortisol and cyclic AMP before and after nine shocks in five subjects. A prominent finding was the rapid stimulation of the ACTH-cortisol production in all subjects, while no consistent changes in FSH, LH, TSH, HGH or cyclic AMP were found.


Antibody-dependent cellular cytotoxicity (ADCC) against Epstein-Barr virus-determined membrane antigens. I. Reactivity in sera from normal persons and from patients with acute infectious mononucleosis. The capacity of antibodies, directed against EBV-determined membrane antigens (MA), to induce antibody-dependent cellular cytotoxicity (ADCC) against EBV superinfected lymphoid cell lines was investigated. Such reactive antibodies were found, at high dilutions, in sera from normal EBV-positive persons but not in sera from EBV-negative persons. Sera from patients with acute infectious mononucleosis (IM), which develop EBV-specific killer T cells, did not induce ADCC. The difference in specificity between ADCC and EBV-specific killer T cells is briefly discussed.


Dose-related sleep disturbances induced by coffee and caffeine. In a 13-night sleep laboratory study, each of 18 normal young adult males twice received 1 cup of warm water, 1-, 2-, and 4-cup equivalents of regular coffee, a 4-cup equivalent of decaffeinated coffee, and a 4-cup equivalent of caffeine. All beverages were administered 30 min before bedtime according to a balanced Latin-square design. Regular coffee produced dose-related changes in most standard electroencephalogram-electrooculogram (EEG-EOG) sleep parameters, and the 4-cup equivalents of regular coffee and caffeine produced equivalent effects. Decaffeinated coffee had no effect. Regular coffee and caffeine caused rapid eye movement (REM) sleep to shift to the early part of the night and stages 3 and 4 sleep to shift to the later part. Coffee also produced dose-related changes in several subjects estimates of sleep characteristics. These results suggest that coffee and caffeine may be used in normal subjects to induce symptoms mimicking those of insomnia. Such a tool should promote further understanding of insomnia.


Probucol: a new cholesterol-lowering drug effective in patients with type II hyperlipoproteinemia. Type II hyperlipoproteinemia or hyperbetalipoproteinemia (B-HLP), a condition with considerable atherogenic potential, is one of the most difficult lipid disorders requiring treatment. Since this abnormality responds minimally to dietary therapy alone, supplemental drug therapy is usually essential. Although the available bile-sequestering resins are effective in B-HLP, these substances are unpalatable and constipating. Since lifelong drug therapy is necessary as an adjunct to diet in the treatment of B-HLP, the ideal drug should be both effective and well tolerated. Probucol, a new cholesterol-lowering drug in tablet form without serious adverse effects, was evaluated in a 12-wk double-blind crossover trial in 11 patients with B-HLP whose serum cholesterol levels were in excess of 275 mg/dl. Probucol, in a dosage of 500 mg twice daily, produced a 10% or greater reduction in serum cholesterol levels in all 11 patients. Serum cholesterol was lowered (p less than 0.01) from 353 to 291 mg/dl in the entire group receiving probucol. There was no significant change (p greater than 0.1) in serum cholesterol (352 mg/dl) during placebo administration. These were no untoward drug effects during the study, and all patients maintained excellent complicance to the schedule of medication. These results indicate that probucol possesses considerable cholesterol-lowering activity and may be a promising new nontoxic therapeutic agent in type II hyperlipoproteinemia.


The influence of orthophosphate on the renal handling of inorganic pyrophosphate in man and dog. 1. The urinary excretion of inorganic pyrophosphate (PPi), a known inhibitor of the growth and aggregation of crystals of calcium phosphate and calcium oxalate, increases after ingestion of orthophosphate (Pi). This effect may contribute to the apparent ability of oral phosphate to reduce the formation of urinary stones in man. This paper is a study of the mechanism by which Pi increases PPi excretion, investigated by renal clearance techniques in man and renal arterial infusion in dogs. PPi in plasma was measured by an isotope-dilution method after ion-exchange chromatography. 2. The mean renal clearance of endogenous PPi in ten men was 7-9 +/- 1-7 (SE) ml/min, and the mean ratio of PPi clearance to creatinine clearance was 0-08 +/- 0-02 (SE). The oral ingestion of Pi increased the urinary excretion and renal clearance of PPi about threefold, without significantly changing its concentration in plasma. 3. In dogs, the infusion of Pi into one renal artery caused a greater increase in urinary PPi from the infused than from the non-infused kidney, an effect that could be accentuated by simultaneous intravenous infusion of PPi. In dogs, only 1-3% of an injected or infused dose of PPi appeared intact in the urine, regardless of whether it was infused into the systemic or renal circulation. 4. These results suggest that Pi has a direct affect on the kidney to increase the excretion of PPi. It is possible that Pi either interferes with tubular reabsorption of PPi, perhaps by competing for a common tubular transport mechanism, or that Pi diminishes the intrarenal hydrolysis of PPi.


Regulation of the human pyruvate dehydrogenase complex. 1. The pyruvate dehydrogenase complex from human heart has been partially purified and shown to be regulated by a phosphorylation-dephosphorylation cycle similar to that previously found for other mammalian tissues. 2. Incubation of the complex with ATP (2 mmol/1) led to its inactivation associated with the concomitant incorporation into the protein of 32P from the terminal phosphate group of the ATP. Pyruvate, ADP, thiamin pyrophosphate and dichloroacetate diminished the rate of inactivation by ATP. 3. Pyruvate dehydrogenase phosphatase from human heart requires Mg2+  for activity and is sensitive to Ca2+ at concentrations of a few mumol/1. Similar ionic requirements of the skeletal muscle phosphatase have been demonstrated in a crude tissue extract. 4. The activity of pyruvate dehydrogenase in human adipose tissue was less than 10% of typical values in rats. This could be due to the high level of dietary fat consumed by humans, which is known to repress the enzyme activity in rats.


The metabolism in vivo and in vitro of plasma low-density lipoprotein from a subject with inherited hypercholesterolaemia. 1. The metabolism in vivo and in vitro of an abnormal low-density lipoprotein (LDL) obtained from a patient with an inherited form of hypercholesterolaemia was compared with that of LDL obtained from a normal subject. 2. The rates of turnover of the apoprotein of the two types of LDL in a normal subject, and their uptake and catabolism by normal lymphocytes in vitro, were similar. 3. It is concluded that the abnormal behaviour of the patient's LDL may not be due to an abnormality in the apoprotein component.


Inverse relationship in Jamaica between plasma high-density lipoprotein cholesterol concentration and coronary-disease risk as predicted by multiple risk-factor status. 1. The relation between plasma high-density lipoprotein (HDL) cholesterol concentration and multiple coronary-risk factor status has been assessed in fifty-two middle-aged clinically healthy men from urban and rural Jamaica. 2. Rural hill-farmers had a superior exercise performance (assessed by the responses to submaximal test exercise), less body fat, and lower fasting levels for plasma total cholesterol, low-density liproprotein (LDL) cholesterol, total triglyceride and blood glucose than urban businessmen. Mean plasma HDL cholesterol was considerably higher in farmers then businessmen. 3. Multilinear regression analysis showed HDL cholesterol concentration to be independently and inversely correlated with plasma triglyceride, LDL cholesterol and diastolic blood pressure and that these relationships applied across the urban and rural sub-groups. There was also some evidence that HDL cholesterol concentration increased with stature. When these factors were taken into account, age, ethnic group, adiposity, weight, exercise performance, smoking history and blood glucose made no further significant contribution to the prediction of HDL cholesterol concentration. 4. Thus plasma HDL cholesterol concentration was highest in those subjects with the lowest coronary-risk as predicted by their multiple risk-factor status, an observation which supported other evidence that coronary-risk is inversely related to plasma HDL concentration. 5. The results raise the possibility that coronary-risk can be more simply estimated from the plasma HDL cholesterol concentration than from a consideration of other major lipid risk factors and blood pressure.


Effect of some nucleotides on the regulation of glycosaminoglycan biosynthesis. The effect of some nucleotides on UDP-glucose dehydrogenase (EC. 1.1.1.22) and UDP-glucose 4'-epimerase (EC 5.1.3.2) extracted from epiphysial-plate cartilage of newborn pigs was investigated. UDP-xylose acts as a co-operative allosteric inhibitor of UDP-glucose dehydrogenase, whereas it does not inhibit UDP-glucose 4'-epimerase activity: the inhibition of UDP-glucose dehydrogenase results in an increase of UDP-galactose synthesis, in agreement with the equilibrium constant of UDP-glucose 4'-epimerase reaction. Because of the presence of UDP-glucose 4'-epimerase activity in the enzyme extract, the addition of UDP-galactose induces an increase in reaction rate of UDP-glucose dehydrogenase. NADH inhibits both UDP-glucose dehydrogenase and UDP-glucose 4'-epimerase activities: in the presence of non-saturating NAD concentrations, NADH acts as a co-operative allosteric inhibitor of both enzymes. The inhibitory effect of NADH seems to be strikingly correlated with the value of NAD/NADH ratio and pH. In any case, the percentage inhibition of UDP-glucose 4'-epimerase, under the same experimental conditions, is always higher than that of UDP-glucose dehydrogenase.


Regulation of active ion transport in the small intestine. The epithelium of the small intestine can both actively absorb and actively secrete electrolytes and water. Secretion can be elicited in vitro by adding cyclic AMP or a stimulator of intestinal mucosal adenylate cyclase (cholera and Escherichia coli enterotoxins, prostaglandins, vasoactive intestinal peptide) or an inhibitor of cyclic AMP phosphodiesterase (theophylline). Cyclic AMP appears to alter intestinal ion transport at two different loci: it inhibits a coupled influx process for Na+ and Cl- at the luminal border, thereby reducing active absorption of NaCl, and it also stimulates the active secretion of anion (or Na+ and anion). A variety of evidence suggests that these two effects of cyclic AMP reside in different types of cells, the former in villus cells and the latter in crypt cells. The latter process is Na+-dependent and is inhibited by low concentrations of ouabain and ethacrynic acid. Active ion absorption in vitro can be enhanced by (1) stimulating Na+-coupled organic solute absorption with glucose, amino acids and possibly also oligo peptides; (2) reducing the HCO3- concentration and/or pH of the serosal bathing solution; and (3) introducing an alpha-adrenergic agonist. Cholera toxin-induced fluid production in vivo can be diminished by the first of these manoeuvres. The in vivo efficacies of the other two have not been evaluated.


The aetiology of diarrhoea in newborn infants. Diarrhoea is a common problem in newborn infants in hospital nurseries. In the past, sporadic diarrhoea was often attributed to dietary indiscretion by the mother, and epidemic diarrhoea was though to be caused by an unknown infectious agent. Techniques with which to locate non-cultivable viruses and untypable enteropathogenic strains of Escherichia coli allow reevaluation of the aetiology of diarrhoea in newborn infants. Preliminary results from Melbourne, Australia, suggest that most diarrhoea in newborn infants is induced by a specific infectious agent. During 1975 the agent most often identified from sporadic and epidemic diarrhoea in hospital nurseries was a reovirus-like particle ("duovirus"). Enterotoxin-producing strains of E. coli were rarely isolated. Future attempts to protect newborn infants from developing diarrhoea must be based on an accurate understanding of the aetiology of this disease.


Implications of recent virological researches. Rotaviruses (duoviruses) can be found in more than half the cases of acute diarrhoea in children up to the age of six or seven. About that age almost everyone has antibodies to them. Second infections occur and may not be as rare as laboratory findings so far suggest. Very young infants sometimes get subclinical disease-the effect of maternal antibody transmitted across the placenta? Very similar viruses, all possessing a common antigen detectable by immunofluorescence, are known to infect and/or cause diarrhoea in children, calves, piglets, mice, foals and monkeys. The calf virus and the human virus both infect piglets; piglet virus infects calves; we don't know whether any of these can infect children. Other mammals probably have similar diarrhoea viruses. An antigen common to all these viruses is probably in the inner capsid layer, and "species-specific" antigens are probably in the outer capsid layer. A precise test for comparing different strains is bably needed. Adenoviruses possibly cause a smaller proportion of cases of diarrhoea. Coronaviruses, well-known as enteric pathogens of pigs and calves, appear also to infect adults and children. 27 nm particles and 22-23 nm particles of density 1.4 (and other particles) can be found in faeces of children with natural diarrhoea and adults with experimental diarrhoea, sometimes in enormous numbers. It is not yet established whether they cause disease. Rotaviruses, animal coronaviruses and "Norwalk" virus attack the disaccharidase-producing epithelium of the small bowel; adenovirus pathology is unknown.A safe attenuated live vaccine strain of the human rotavirus urgently needs to be developed.


Pathogenic rotaviruses isolated from pigs and calves. Rotavirus is commonly isolated from diarrhoeic calves and pigs. Bacterium-free faecal filtrates containing rotavirus from five different outbreaks of disease in calves all caused diarrhoea and clinical illness in gnotobiotic calves and five different isolates from pigs were inoculated into gnotobiotic pigs with similar results. The author was unsuccessful in finding an avirulent strain although one of the calf isolates was from a non-diarrhoeic calf. The laboratory strain of calf virus retained its virulence after being passaged seven times in gnotobiotic calves, which included sucrose density gradient purification on two occasions. The calf tissue culture-adapted virus retained its virulence. Rotavirus isolates from humans, calves, pigs and foals were infectious to pigs. Although sharing a common antigen the viruses were separable according to host infectivity, virulence and neutralizing antigens. In both calves and pigs the main lesion was loss of the epithelial cell of the small intestine and stunting of villi. Passive protection of the calf and pig was poor. Circulating antibody was not protective and although high levels of clolostral antibody in the gut lumen at the time of infection protected calves clinically, the antibody level secreted in milk declined 10-fold 48 hours after parturition. Frequently other viruses are found together with rotavirus in cases of diarrhoea. Their role is being investigated.


Recent advances in the aetiology of viral gastroenteritis. Studies with the human reovirus-like (HRVL) agnet, also designated rotavirus and duovirus, have revealed that it is a major aetiological agent of diarrhoea of infants and young children in many parts of the world. In a study of patients admitted with a diarrhoeal illness to the Children's Hospital of the District of Columbia in the United States from January 1974 to June 1975, it was found that half of the patients studied by both virus shedding (by electron microscopy) and serological (complement-fixation) techniques demonstrated evidence of infection with the HRVL agent. The temporal distribution of infections with the HRVL agent followed a seasonal pattern with this agent being shed exclusively by patients admitted during the cooler months of the year. Electron microscopic examination of stools was as efficient as serological methods for detecting infection with the HRVL agent. We also initiated studies to determine the possible mode of transmission of the HRVL agent by studying contacts of hospitalized patients. We found that 35% parents of patients with HRVL infections were also infected with the HRVL agent. Serological studies revealed that the HRVL agent was antigenically related to the Nebraska Calf Diarrhoea Virus, the epizootic diarrhoea of infant mice virus, the SA-11 virus, and the "O" agent.


The activation of adenylate cyclase by cholera toxin: possible interaction with the nucleotide regulatory site. The application of cholera toxin to intact cells causes a stimulation of adenylate cyclase activity. The effect is characterized by a lag period followed by a progressive rise in enzyme activity over several hours. Only a few minutes' exposure to the toxin is required to produce effects lasting over several days. Stimulation of adenylate cyclase by cholera toxin in broken cell preparations requires the presence of nicotinamide-adenine dinucleotide (NAD) and an unidentified component of the cytosol. Guanyl nucleotides and certain non-hydrolysable analogues of guanosine triphosphate also stimulate adenylate cyclase. Stimulation by the analogues results in a highly activated enzyme which has characterisitcs similar to those of adenylate cyclase after stimulation by cholera toxin. Thus the stimulation is irreversible, the enzyme may be "solubilized" by non-ionic detergents in the activated state, and responses to certain hormones are enhanced. Therefore the possibility exists that cholera toxin acts on the guanyl nucleotide regulatory protein of the adenylate cyclase complex. In exploring this possibility it was found pretreatment with cholera toxin not only blocked the stimulatory effect of subsequently added guanylylimidodi-phosphate (GppNHp) but that the latter reduced the stimulation by toxin. Similarly, pretreatment with GppNHp blocked the effect of cholera toxin. The similarities in the effects of cholera toxin and GppNHp, together with the mutual interference of their activities, suggests that cholera toxin acts at the same regulatory site at which guanyl nucleotides exert their effects on adenylate cyclase.


Behavioral manifestations of the Kleine-Levin syndrome. The authors present a case of the Kleine-Levin syndrome, and review the literature to assess the various ways this illness can appear. Psychiatric manifestations not only obscure the diagnosis, but may lead to inappropriate treatment.


Granular-cell myoblastoma of the cercum: report of a case. A 9-mm granular-cell myoblastoma of the cecum found incidentally during appendectomy in a 17-year-old-girl is reported. Electronmicroscopic findings favor origin of the granular cells from an undifferentiated mesenchymal (fibroblast-like) cell. Review of the small number of previously reported cases revealed three involving the cecum, one each in the ascending and transverse colon and two in the rectum. Four patients were asymptomatic and their lesions were found incidentally. Three lesions simulated maligancy clinically; these patients underwent right hemicolectomy.


Carcinoma of anal-ductal origin: report of a case. A 57-year-old man who had a carcinoma proven to be of anal ductal origin is presented. Recurrent perianal fistulas were treated locally for two years until biopsy proved the malignancy. Gross and histologic examination of the specimen removed by abdominoperineal resection demonstrated the anal ductal origin of this tumor. Recurrence was seen in less than a year.


[The neuroendocrinology of sleep].

[The neuroendocrinology of sleep]. It is now well established that some pituitary hormones have a pattern of secretion closely linked to the sleep-waking cycle. These data open a new approach to the problem of sleep disturbances and suppose a completely new evaluation of their meaning.


[The effects of calcitonin, glucagon and the dibutyryl derivative of cyclic AMP on plasma-renin activity (author's transl)]. Infusion of 2 MRC/kg X h calcitonin in anaesthetised dogs produced a significant increase in plasma-renin activity, clearance of 51Dr-EDTA and 125I-o-iodohipuric acid, heart rate and urinary excretion of sodium, potassium, calcium and phosphates, while serum electrolytes and mean arterial pressure markedly fell. Infusion of 5 mug/kg X min glucagon produced a significant fall of plasma-renin, heart rate rose, but arterial mean pressure fell, and serum and urinary electrolytes did not change significantly, Cyclic AMP (dibutyryl-cAMP) significantly stimulated renin at a dose of 5 mg/min, while there were no significant changes in blood pressure, heart rate and serum and urinary electrolytes.


[Idiopathic hypophosphataemic osteomalacia (author's transl)]. In an adult with sporadic idiopathic osteomalacia an increased phosphate clearance, hypophosphataemia, normocalcaemia, normal serum-25-hydroxycalciferol and an only slightly increased immunoreactive parathormone were found. Intestinal 47Ca absorption was clearly decreased. Radiologically and histologically there was a clear-cut defect of skeletal mineralisation. Under treatment with daily doses of 1-1.25 mg of vitamin D3 the 25-hydroxycalciferol level increased markedly, the immunoreactive parathormone decreased slightly. Serum calcium and hypophosphataemia remained unchanged and intestinal 47Ca absorption was improved. Already 4 weeks after commencing treatment pain and defective gait of the patient disappeared. Radiologically skeletal changes were improved after 7 months. However, histologically no significant bone healing had occurred. The biochemical findings of this disease correspond to those of familial hypophosphataemic (vitamin-D-resistant) rickets. The therapeutic effects of pharmacological doses of vitamin D resemble those in pseudo-vitamin-D-deficient rickets. The pathogenesis of idiopathic osteomalacia of the adult remains unclear. Vitamin D metabolism is unchanged as far as the stage of 25-hydroxycholecalciferol. It is unknown if a disorder of the renal synthesis of 1,25-dihydroxycholecalciferol or a peripheral resistance to the effects of this metabolite exists. In addition a defect of the tubular phosphate reabsorption independent of parathormone and vitamin D is assumed.


Hormone-induced dispersion or aggregation of carotenoid-containing smooth endoplasmic reticulum in cultured xanthophores from the goldfish, Carrassius auratus L. A novel organelle movement is described in goldfish xanthophores. Smooth endoplasmic reticulum, containing carotenoids pigments, undergo MSH or c-AMP-induced dispersion. This dispersion is independent of RNA and protein synthesis, not modulated by c-GMP and inhibited by cytochalasin B but only at the relatively high concentration of 10 mug/ml. Epinephrine induces aggregation, a process that is inhibited by colchicine. These results suggest, but do not prove, the involvement of microfilaments in dispersion and microtubules in the aggregation of carotenoid-containing endoplasmic reticulum.


Regulation of lactogen specific binding sites in rat liver: studies on the role of lactogens and estrogen. Regulation of the lactogen specific binding sites of rat liver was studied in several different high lactogen states. The specific binding of [125I]iodo-hGH was determined as a measure of these sites. Hypophysectomy of pregnant rats produced a much smaller decrease in hepatic binding of [125I]iodo-hGH than was seen in nonpregnant females. Three different prolactin secreting tumors (MtT/F4, MtT/F45, MtT/W5) produced greatly elevated levels of both rPRL and hepatic binding of [125I]iodo-hGH in both male and female rats. The increased binding reflected an increase in the concentration of lactogen specific membrane binding sites. There was no change in the apparent affinity of binding. Hypophysectomy of tumor-bearing rats was not followed by a decrease in the concentration of hepatic sites. Pituitary transplants beneath the kidney capsules of hypophysectomized (hypox) rats produced elevated rPRL levels and increased [125I]iod-GH specific binding to hepatic membranes. The elevation of serum rPRL preceded the increase in hepatic binding. There was a direct correlation between rPRL levels attained by 8 days after implantation and the level of hepatic binding. Estrogen treatment did not alter this correlation. Hormonal replacement with prolactin in combination with various hormones failed to induce the lactogen specific hepatic sites in estrogen treated hypox males, and did not prevent the marked decrease in hepatic binding observed in females following hypophysectomy. It is suggested that chronic elevation of prolactin plays a role in inducing lactogen specific binding sites in rat liver. Estrogen's inductive action seems to be largely effected at the pituitary level.


Inhibitory and facilitatory areas of the dorsal medulla mediating ACTH release in the cat. To define the role of the dorsal medulla in the control of release of ACTH, the authors stimulated electrically (30 sec, 100 muA, 50 Hz) 50 sites in the vicinity of the solitary nuclei of 11 cats anesthetized with chloralose/urethane. Responses of arterial pressure to electrical stimulation were not correlated significantly with release of ACTH. Indirect effects of changes in arterial pressure could not explain changes in release of ACTH. Concentrations of ACTH were measured by radioimmunoassay. Active areas associated with the solitary nucleus were : 1) lateral inhibitory: ventral and lateral to the solitary tract (mean delta ACTH:-153, -86, -97 pg/ml at 1.5, 3.0 and 6.0 min respectively; P less than 0.01); 2) medial inhibitory: medial dorsal motor nucleus of the vagus and extending to the midline (mean delta ACTH: -81, -107, -67 pg/ml; P less than 0.01); and 3) intermediate facilitatory: lateral nucleus intercalatus and adjacent reticular formation (mean delta ACTH: +105, +158, +4 pg/ml; P less than 0.01). The former two areas contain neurons activated by atrial stretch, and the latter area contains neurons inhibited by atrial stretch. Since changes in ACTH levels are inversely correlated with atrial stretch, the results suggest that the changes in release of ACTH are the result of direct stimulation of neural systems of the solitary nuclei mediating release of ACTH in response to hemodynamic changes.


Inhibitory and facilitatory areas of the rostral pons mediating ACTH release in the cat. To define the role of the rostral pons in the control of release of ACTH, we stimulated electrically (30 sec, 200 muA, 50 Hz) 128 sites in the dorsal rostral pons of 20 cats anesthetized with chloralose/urethane. Responses of arterial pressure to electrical stimulation were prevented by lesions placed previously in the medulla. Plasma concentrations of ACTH were measured by radioimmunoassay. Active areas consisted of three regions: 1) lateral inhibitory: Locus subcoeruleus and anteroventral locus coeruleus (mean deltaACTH: -189, -164, -145 pg/ml at 1.5,3.0 and 6.0 min respectively, P less than 0.01);2) intermediate facilitatory:principal locus coeruleus and lateral ventral tegmental nucleus (mean deltaACTH: +81, +68, +37 pg/ml; P less than 0.05); and 3) medial inhibitory: dorsal tegmental nucleus, dorsal raphé and medial ventral tegmental nucleus (mean deltaACTH; -211, -212, -115 pg/ml; P less than 0.01). The former two areas received direct projections from medullary neurons activated or inhibited by atrial stretch, and, in turn, give rise to adrenergic and cholinergic projections to the medial hypothalamus. Since the release of ACTH is inversely correlated with right atrial stretch, the results suggest that the lateral inhibitory area and the intermediate facilitatory area are involved in mediation of changes in release of ACTH in response to hemodynamic changes.


Concentration of cyclic AMP in rat liver as a function of the insulin/glucagon ratio in blood under standardized physiological conditions. The concentration of insulin and glucagon in peripheral blood and the concentration of cAMP in liver was followed in rats throughout a 48 hour starvation period and up to 6 hours afer refeeding glucose or casein. By so changing the insulin/glucagon molar ratio from minimum to maximum values, simultaneous inverse changes in the concentration of hepatic cAMP could be induced. The study, thus, suggests that during a starvation-refeeding cycle the level of cAMP in the liver is regulated predominantly by the insulin/glucagon ratio in the blood. Possible criticisms of this conclusion are discussed.


Studies on the mechanism of the antilipolytic effects of growth hormone. Ovine growth hormone (1 mug/ml) antagonized the lipolytic action of epinephrine (0.25 mug/ml) in segments of adipose tissue obtained from hypophysectomized rats, but a lag period of about 10 min was required. When added simultaneously with epinephrine, growth hormone neither reduced the maximal accumulation of cyclic AMP which occurred at 3 min nor accelerated the return to basal levels. Only when tissues were exposed to epinephrine 15 min after preincubation with growth hormone was cyclic AMP accumulation compromised. Growth hormone also produced a delayed increase of about 20% in the activity of a low Km cyclic nucleotide phosphodiesterase, which might have contributed to the decrease in cyclic AMP accumulation. The increase in phosphodiesterase activity probably did not account for the antilipolytic effect, however, since antilipolysis was evident before the increase in phosphodiesterase activity could be  detected. The antilipolytic effects of growth hormone similarly could not be attributed to the decrease in cyclic AMP concentrations, for when added simultaneously with epinephrine the antilipolytic effects did not occur until after the evanescent changes in cyclic AMP had passed. Growth hormone added simultaneously with epinephrine or 30 min later significantly decreased the activity of protein kinase assayed in the absence of exogenous cyclic AMP, but did not change total protein kinase activity as measured in the presence of a saturating concentration of cyclic AMP. This effect of growth hormone was evident as early as 3 min after addition of the hormone and may at least partially account for the antilipolytic effect.


Studies on oxidative phosphorylation and steroidogenesis by ovarian mitochondria after gonadotropic stimulation. No differences in oxidative phosphorylation or in the per cent of [4-14C]progesterone were found in ovarian mitochondria of immature rats after treatment with 20 IU of pregnant mare serum gonadotropin (PMSG) iv 30 min before killing. However, treatment of immature rats with 20 IU of PMSG sc 54 h prior to killing decreased the ADP:O ratio and increased the per cent of [4-14C]cholesterol conversion. Electron microscopic studies showed that mitochondria with lamellar cristae were prominent in ovaries of untreated rats, while large pleomorphic mitochondria and mitochondria with tubulovesicular cristae dominated in ovaries of PMSG-treated rats. Ovarian homogenates separated by zonal centrifugation showed three peaks od cytochrome oxidase activity which shifted to the heavier end of the gradient after PMSG treatment. These studies suggest that PMSG treatment influences ovarian mitochondria, possibly by stimulating the synthesis of additional functional components and/or the biogenesis of new mitochondria. Aminoglutethimide addition to bovine luteal mitochondria decreased steroidogenesis by 60% when succinate was used as substrate. However, there was a 16% increase in the ADP:O ratio, apparently due to a decrease in oxygen utilization. When oligomycin was added to luteal mitochondria, there was a 30% decrease in the ACP:O ratio but a 300% increase in [4-14C]cholesterol conversion. Dinitrophenol also decreased mitochondrial steroidogenesis. These results suggest that energy obtained from succinate oxidation can be diverted from phosphorylation to support steroidogenesis.


On the possible role of thiol groups in the insulin-releasing action of mercurials, organic disulfides, alkylating agents, and sulfonylureas. The thiol activity of pancreatic islets was spectrophotometrically assayed as the formation of 6-mercaptonicotinic acid from the organic disulfide, 6,6'-dithiodinicotinic acid. Islets containing more than 90% beta-cells were microdissected from non-inbred ob/ob-mice. Comparisons of intact with homogenized islets indicated that the organic disulfide penetrates relatively slowly into the beta-cells. When tested at concentrations know to enhance insulin release, p-chloromercuribenzene-sulfonic acid almost completely blocked the thiol activity of intact islets, whereas no significant effect was observed with iodoacetamide, D-glucose, or glibenclamide. Although glibenclamide had no demonstrable effect on the thiol activity of free L-cysteine, the binding of glibenclamide to serum albumin was decreased by blocking the albumin thiols with azobenzene-2-sulfenyl bromide. The uptake of glibenclamide by pancreatic islets was inhibited by cysteine or reduced glutathione. Cysteine, as well as 6,6'-dithiodinicotinic acid, also seemed to interact negatively with glibenat organic mercurials and disulfides stimulate insulin release by blocking thiol groups in the beta-cell plasma membranes. The thiol groups involved in iodoacetamide-induced secretion may escape detection by the assay employed, or target groups other than thiols may be involved. The data on glibenclamide are compatible with, but do not unequivocally support, the notion that thiol groups may play a role in sulfonylurea-induced insulin release.


Effects of ACTH on plasma levels of adrenal steroids in essential hypertension. Plasma concentrations of progesterone (P), deoxycorticosterone (DOC), 17-hydroxyprogesterone (17-OH P), corticosterone (B), deoxycortisol (S), cortisol (F) and aldosterone (A) in 8 control subjects (mean age: 40.5 years) and 10 patients with essential hypertension (EH) (mean age: 48.5 years) were determined before, 4 and 8 hours after an infusion of ACTH at a rate of 25 units per 8 hours. Secretion rates (SR) of 18-hydroxy-11-deoxycorticosterone (18-OH DOC) were measured 24 hours before and again on the day of ACTH infusion. All subjects were studied on the fourth day of a diet containing 135 mEq of sodium and 90 mEq of potassium. There was no statistically significant difference between 8 control subjects and 10 patients with EH in the 7 plasma steroid levels and the SR of 18-OH DOC before ACTH infusion. The mean plasma P response to ACTH was slightly lower in controls than in patients with EH, while that of 17-OH P (in male subjects) was slightly higher. The mean plasma B response was significantly lower after 4 hours of ACTH infusion (p less than 0.01), while that of DOC was significantly higher after 8 hours of ACTH infusion (p less than 0.05) in patients with EH. The mean plasma S rose significantly more in patients with EH (p less than 0.025) at 4 and 8 hours after ACTH infusion. The mean plasma F response to ACTH infusion was slightly lower in patients with EH than in controls. The mean response of 18-OH DOC SR to ACTH infusion was slightly higher in patients with EH than in controls. The mean plasma A response was significantly higher in patients with EH than in controls 4 (p less than 0.05) and 8 hour (p less than 0.001) after an ACTH infusion. These results could be explained in part by abnormalities in the 17- and 11-hydroxylase systems, and that the abnormality in 11-hydroxylation was more pronounced than that in the 17-position. Furthermore, we suspect that the sensitivity of adrenal aldosterone to ACTH might be increased or another accelerated pathway to aldosterone biosynthesis might exist in patients with EH.


Plasma immunoassayable ACTH levels during and after hydrocortisone infusion in patients with Cushing's disease. The plasma ACTH responses to hydrocortisone infusion were compared in patients with Cushing's disease and primary adrenocortical insufficiency. In 4 patients with primary adrenocortical insufficiency, plasma ACTH levels were suppressed exponentially after administration of a relatively large dose of hydrocortisone (1.0 mg/kg/1.5 hr-3.0 mg/kg/2 hr). In patients with post-adrenalectomized Cushing's disease (4, bilateral; 1, unilateral), plasma ACTH suppression was delayed. Plasma ACTH levels, expressed as a percentage of the basal concentrations, were significantly less suppressed in patients with Cushing's disease than in patients with primary adrenocortical insufficiency 90 (p less than 0.05) and 120 (p less than 0.05) min after the beginning of infusion. When 0.5 mg/kg hydrocortisone was infused over a period of 1.5 hr, suppression was also delayed in Cushing's disease, and plasma ACTH levels were less suppressed in 4 patients with Cushing's disease than in 4 patients with primary adrenocortical insufficiency at 30 (p greater than 0.05), 45 (p greater than 0.05) 60 (p less than 0.05) min.


Enzymatic radioiodination of succinyl cyclic AMP tyrosine methyl ester by lactoperoxidase and radioimmunoassay for cyclic AMP. A highly specific and simple radioimmunoassay for cyclic AMP with a sensitivity of 0.04 picomoles/tube has been developed according to the method of Steiner et al., using 125I-succinyl cyclic AMP tyrosine methyl ester as a tracer. The tracer with higher immunoactivities could be simply and constantly prepared by an enzymatic iodination procedure utilizing lactoperoxidase, radioactive iodide and hydrogen peroxide generated by glucose-glucose oxidase system, rather than by chloramine-T procedure.


Effects of lipolytic and antilipolytic drugs on metabolism of adenosine 3':5'-monophosphate in brown adipose tissue of cold acclimated rats. The intracellular level of adenosine 3':5'-monophosphate (cyclic AMP) and its stimulation in vitro by norepinephrine were studied in brown and white adipocytes from rats adapted to constant or fluctuating cold. Cold acclimatization had no effect on the basal cyclic AMP intracellular content in both tissues, but the level in brown adipocytes was four-fold higher than in the white ones. Addition of norepinephrine in the incubation medium doubled the cyclic AMP content of white adipocytes from control or fluctuating-cold-adapted rats, and enhanced four-fold in constant-cold-adapted rats. In brown adipocytes norepinephrine increased cyclic AMP levels in the first two groups, but had no effects in constant-cold-adapted rats. In the two tissues of control and fluctuating-cold-adapted rats the norepinephrine action was increased by phentolamine and decreased by propranolol. The lack of response to norepinephrine of brown adipocytes from constant cold-adapted rats was not due to the predominance of the alpha component of hormone receptors. Antilipolytic drugs (nicotinic acid, insulin and prostaglandin E2) inhibited the action of norepinephrine on white adipocytes; only prostaglandin E2 had an effect on brown ones.


Nuclear-magnetic-relaxation studies of the interaction of inhibitor with the threonine-sensitive aspartokinase of Escherichia coli. The nature of the feedback inhibition of the bifunctional enzyme, aspartokinase I-homoserine dehydrogenase I of Escherichia coli was studied using 13C nuclear magnetic resonance (NMR). Since aspartokinase is activated by Mn(II), the interaction of the inhibitor L-threonine (specifically enriched to 90% 13C in the carboxyl carbon) with the metal-enzyme complex was studied. Spin-lattice (T1) and spin-spin (T2) relaxation times were determined by the partially relaxed Fourier transform method and line-width measurements respectively at 20 MHz. The pronounced broadening of the DL-threonine carboxyl peak in the presence of the Mn(II)-enzyme complex indicates that an L-threonine binding site is close to the metal binding site of the kinase active site. The non-identity of (T1)*M and (T2)*M indicates that conditions of fast exchange prevail. The (T1)*M/(T2)*M ratio was used to estimate a correlation time of 2.0 ns for the dipolar interaction at 25 degrees C. An estimate for the distance between Mn(II) and the threonine carboxyl carbon of 4.4 A (0.44 nm) was obtained. This 13C NMR study has thus located one of the two classes of threonine regulatory sites which exist per subunit; the threonine site identified here is at the aspartokinase active site, adjacent to the catalytic metal site.


Uridine--cytidine kinase from foetal and adult rat liver. Purification and study of some properties. Partial purification of uridine--cytidine kinase (EC 2.7.1.48) from foetal rat liver by chromatography on DEAE-cellulose gives two active fractions. The first in order of elution was identified as a form specific for foetal liver. It was purified 300-fold. The second fraction was common to foetal, and adult rat liver and spleen and was purified 20-fold. The foetal fraction of the enzyme was found to be heat-sensitive and protected against inactivation by PO34- anions. The two isolated forms have different apparent Km for uridine, respectively 410 muM for the foetal form and 52 muM for the adult form.


Kinetic transients in the reduction of aldehydes catalysed by liver alcohol dehydrogenase. The transient-state kinetics of enzymic reduction of acetaldehyde and benzaldehyde by NADH, catalyzed by horse liver alcohol dehydrogenase, have been examined under single-turnover conditions, obtained by carrying out reactions either with limiting amounts of enzyme in the presence of 20 mM pyrazole or with limiting amounts of substrate. Analysis of the variation with substrate, coenzyme, and enzyme concentrations of amplitudes and time constants for the exponential transients observed at 328 nm and 300 nm shows that the kinetics of enzymic aldehyde reduction are qualitatively and quantitatively consistent with the relationships derived in the preceding paper for an ordered ternary-complex mechanism involving identical and independent catalytic sites. It is concluded that there is no evidence whatsoever for the kinetic significance of a half-of-the-sites reactivity or any other kind of subunit interaction in the liver alcohol dehydrogenase system. The biphasic transients observed at 328 nm for the reduction of aromatic aldehydes such as benzaldehyde are a normal kinetic characteristic of the ordered ternary-complex mechanism, being attributable to accumulation of the ternary enzyme-NAD-product complex when product dissociation from this complex is slow in comparison to its formation by ternary-complex interconversion.


An improved procedure for automated Edman degradation used for determination of the N-terminal amino acid sequence of human transcobalamin I and human intrinsic factor. An improved procedure for automated Edman degradation is presented. Three programs are described, one with double cleavage and two with single cleavage. The programs presented are characterized by a reversed delivery scheme for buffer and phenyl isothiocyanate, and by reduced cleavage times. The modified procedures applied on automated Edman degradation of the vitamin B12-binding proteins human transcobalamin I and human intrinsic factor, containing approximately 390 and 350 amino residues respectively, gave the following N-terminal amino acid sequences: Human transcobalamin I Glu-Ile-Cys-Glu-Val-Ser-Glu-Glu-Asn-Tyr-Ile-Arg-Leu-Lys-Pro-Leu-Leu-Asn-Thr-Met-Ile-Gln-Ser-Asn-Tyr-Asn-?-Gly- Human intrinsic factor Ser-Thr-Gln-Thr-Gln-Ser-Ser-Cys-Ser-Val-Pro-Ser-Ala-Gln-Glu-Pro-Leu-Val-Asn-Gly-Ile-Gln-?-Leu-Met-Glu-Thr- The background accumulation seems to be related not only to the length of the polypeptide chain being degraded, but also to the content of serine (and possibly threonine). A possible N leads to O acyl shift during the cleavage is a tentative explanation. The programs here represented lead to a significant reduction in background compared to conventional programs and allowed considerable prolongation of the degradations.


Effect of cerebral injury on cerebrospinal fluid cyclic AMP concentration. Cerbrospinal fluid (CSF) cyclic adenosine-3', 5'-monophosphate (cAMP) was measured in rabbits after experimental brain injury as well as in patients with cerebral contusion and cerebral concussion. In rabbits a marked elevation lasting for two weeks was observed. From the third week onwards after the injury the CSF cAMP concentration was lower than the basal level before the injury. Dexamethasone partly inhibited the elevation. Ethanol treatment decreased the CSF cAMP values. In man, the CSF cAMP concentration was significantly higher in patients with cerebral contusion than in those with cerebral concussion. Also in the latter group the cAMP values were higher than in control patients. Due to the clear differences between the various groups, measurement of cAMP concentration in CSF might have diagnostic value in evaluation of the severity of cerebral lesion in the acute phase. Also the activities of some enzymes were measured, and the results were parallel with cAMP changes but less striking.


Is insomnia a disease of slow-wave sleep? The aim of this paper is to examine if a disturbance of slow-wave sleep may partly account for the imbalance between waking and sleep observed in insomnia. 40 normal subjects and 40 insomniacs were recorded in the laboratory. No direct interregulation appeared between total sleep and REM sleep on one hand, and between slow-wave sleep and REM sleep on the other. Slow-wave sleep, however, was linked to the waking-sleep imbalance, as low values of stages 3 and 4 were statistically associated with low total sleep duration. The reduction of slow-wave sleep could not merely be attributed to an increased pressure of wakefulness. Our results indicate that it represents probably a disturbance in itself, perhaps related in some cases to a precocious senescence of sleep, but do not account alone for all sleep disturbances.


Stimulation of lymphocytes by allogeneic lymphocytes and lymphoblasts in the presence of anti-HLA antisera. Anti-HLA alloantisera inhibit mixed lymphocyte responses in which normal lymphocytes are used as stimulator cells. These same antisera are unable to inhibit lymphocyte proliferative responses stimulated by lymphoblastoid cells from cultured lymphoid cell lines. They also fail to inhibit either the generation of cytotoxic effector cells by lymphoblastoid cells or lymphocyte-mediated cytotoxicity against the lymphoblasts. Although the number of HLA antigens on the surface of lymphoblasts is reported to be greater than on normal lymphocytes, the failure of alloantisera to inhibit lymphoblast-induced responses in vitro does not appear to be due to insufficient amounts of antiserum to react with the antigenic sites. Rather, the data are interpreted to suggest that antigens which are not HLA and are not closely associated with HLA on the lymphocyte membrane are responsible for the stimulation of allogeneic lymphocytes by lymphoblastoid cells. Although lymphoid cell lines are known to contain the genome of the Epstein-Barr virus, antisera against products of the viral genome fail to inhibit proliferative responses to lymphoblastoid cells, suggesting that these antigens do not directly participate in lymphocyte activation.


An ultrastructural study of two different responses of mouse mast cells to transplantation antibodies directed against the same transplantation antigens. The present paper describes an ultrastructural study of two different kinds of behavior of mouse mast cells during two immunological reactions induced by transplantation antibodies: the direct allogeneic anaphylactic degranulation and the serocytotoxicity. In both situations, the same alloantigens, born by the mast cells themselves, are the targets of the reaction, but the first one is mediated by anaphylactic alloantibodies whereas the second one is mediated by cytotoxic alloantibodies in the presence of complement. The comparison of the ultrastructural aspects of the cells in these two systems demonstrated that mast cells can behave in two different ways depending on the nature of the immunological agents utilized. First, a physiological degranulation process with active granule expulsion was observed. This process was shown to be identical to the one induced in classical in vitro anaphylaxis or by histamine releasers such as compound 48/80. A second type of behavior was a lethal, complement-dependent, cell lysis without active granule expulsion.


Renal adenylate cyclase-effects of diuretics. The in vitro effect of various diuretics on rat kidney adenylate cyclase was investigated in crude homogenates of the cortex, the outer and inner medulla. 10-3 M furosemide inhibited adenylate cyclase by 40% in the cortex, by 16% in the outer medulla and by 43% in the inner medulla. 10-3 M ethacrynic acid inhibited adenylate cyclase activity by 65% in the cortex, 59% in the outer medulla and by 57% in the inner medulla. Amiloride produced no significant inhibition of the adenylate cyclase reaction. In the cortex, furosemide partially inhibited adenylate cyclase under basal, fluoride-stimulated and parathyroid hormone-stimulated conditions. Ethacrynic acid produced a strong inhibition of adenylate cyclase activation by F- and parathyroid hormone. In the inner medulla 10-2 M F- and 1 mU antidiuretic hormone reversed the furosemide effect on adenylate cyclase. Ethacrynic acid produced a strong inhibition of adenylate cyclase in the presence of F- and antidiuretic hormone. It is suggested that inhibition of renal adenylate cyclase might be a possible mode of action of certain diuretics.


Preferential blockade of presynaptic alpha-adrenoceptors by yohimbine. Strips of the rabbit main pulmonary artery were preincubated with 3H-noradrenaline. 3 X 10(-8) -- 10(6) M yohimbine enhanced the overflow of tritium and the smooth muscle contraction induced by transmural sympathetic nerve stimulation. The increase of the stimulation-evoked overflow of tritium was prevented by a high concentration of oxymetazoline. The results indicate that yohimbine is more potent in blocking the presynaptic than the postsynaptic alpha-adrenoceptors of the artery.


Antagonism by Ca2+ of the inhibitory action of PGE1 on the electrically induced vasoconstriction in the rabbit ear artery. The inhibitory action of PGE1 on vasoconstrictor responses to nerve stimulation of isolated ear arteries of rabbits was studied in the presence of 1.0 to 7.5 mM external calcium concentration. Raising the [Ca]0 in the medium increased the electrically induced vasoconstriction and antagonized the inhibitory effect of PGE1 on constrictor responses. This antagonism is probably due to an interaction between Ca2+ and PGE1 in the sympathetic nerves to the vessel.


Inhibition by alpha- and beta-adrenoceptors of the twitch response to transmural stimulation in the guinea-pig vas deferens. Noradrenaline as well as the indirectly acting amines tyramine and phenethylamine either enhance or inhibit the twitch response of the transmurally stimulated, isolated guine-pig vas deferens, thus partly confirming previous reports. In both cases enhancement is annulled by alpha-adrenoceptor blockers. The twitch inhibition caused by noradrenaline is abolished by alpha- + beta2-adrenoceptor blockers, but not by either blocker alone. The inhibition caused by the indirectly acting amines is largely abolished by alpha-adrenoceptor blockers. Clonidine strongly inhibits the twitch. This effect if promptly removed by phentolamine. After blockade of the neurally induced twitch by tetrodotoxin, noradrenaline and the indirectly acting amines have no effect or slightly enhance the twitch elicited by transmural stimulation of the smooth muscle. It is concluded that exogenous noradrenaline acts on postjunctional stimulatory alpha-adrenoceptors and on inhibitory alpha- and beta2-adrenoceptors, which are presumably prejunctional. In the unstimulated preparation contracted by acetylcholine, noradrenaline causes further contraction which is changed into relaxation after phentolamine. This relaxation is abolished by butoxamine, suggesting that noradrenaline may also act on inhibitory postjunctional beta2-adrenoceptors. The twitch-inhibiting effect of endogenous noradrenaline, released by nerve stimulation or by indirectly acting amines, appears to be primarily mediated by prejunctional alpha-adrenoceptors.


Effect of presynaptic alpha-adrenoceptor blockade on responses to cardiac nerve stimulation in anesthetized dogs. Phentolamine caused significant potentiation of chronotropic responses to cardioaccelerator nerve stimulation (CANS) in pentobarbital-anesthetized dogs without impairing neuronal reuptake of norepinephrine. In a separate group of dogs, desipramine produced slight augmentation of CANS responses which were potentiated further by phentolamine administration. These results provide support for the hypothesis that presynaptic alpha-adrenergic receptors play an inhibitory role in sympathetic neurotransmission and demonstrate that this mechanism is also functioning during stimulation of the cardioaccelerator nerve in pentobarbital-anesthetized dogs.


Variations in the number of uterine angiotensin receptors following changes in plasma angiotensin levels. 3H-labelled angiotensin II binding to receptor sites was studied in plasma membranes isolated from myometrial homogenates of uterine horns. Removal of the kidneys, which results in the disappearance of plasma angiotensin II, was followed 19 h after nephrectomy by an increase in the number of uterine receptor sites without significant variation in the apparent dissociation constant. Acute pressor i.v. injection of angiotensin II into nephrectomized rats immediately before removing uteri, did not affect the number of uterine angiotensin receptors, whereas long-lasting angiotensin infusion did reduce the number of receptors. These changes cannot be accounted for by variations in the occupancy of receptor sites. These results demonstrate that the number of angiotensin receptors, at least in uterine contractile cells, is affected by chronic variations of endogenous angiotensin levels. The relation between the specific supersensitivity to angiotensin II observed in uteri from nephrectomized rats and the variations at the receptor level is discussed.


Neuromuscular blocking action of cadmium and manganese in isolated frog striated muscles. In isolated frog sciatic nerve--gastrocnemius muscle preparations, exposure for 60 min to Cd2+ (0.1-2 mM) caused a marked attenuation of the twitch tension developed by stimulation of nerves but no or only a slight inhibition of the tension produced by direct stimulation at maximum twitch height. The inhibitory effect was partially reversed by 1 mM cysteine. In frog sartorius muscles, the addition of Cd2+ (0.1 mM) alos abolished the end-plate potential evoked by nerve stimulation but did not suppress potential changes induced by iontophoretically applied acetylcholine. The addition of Mn2+ (2 and 5 mM) attenuated the response of muscles to both direct and indirect stimulation; a greater attenuation of the latter was observed. The inhibition was not reserved by cysteine but was partially reversed by excess Ca2+. Contractile responses of frog rectus abdominis muscles to acetylcholine were not significantly affected by Cd2+ and Mn2+ but were attenuated by d-tubocurarine in a dose-dependent manner. Impulse conduction along sciatic nerves was not impaired by Cd2+ and Mn2+ but was by procaine. It appears that Cd2+ interferes with the release of acetylcholine from motor nerve terminals by reducing the transmembrane influxes of Ca2+, the influx possibly relating to SH groups of membrane constituents.


Selective activation of nicotinic and muscarinic transmission in cardiac sympathetic ganglion of the dog. The myocardial contractile responses to electrical stimulation of different segments of the thoracic sympathetic trunk in the pentobarbital-anesthetized dog were studied. Electrical stimulation of the sympathetic trunk between the second and third thoracic ramus and caudal limb of the ansa subclavia elicited myocardial contractile responses which were not completely antagonized by the ganglionic blocking agent chlorisondamine. Atropine in addition to chlone had no effect. When the sympathetic trunk between the third and fourth ramus was electrically stimulated, the response was partially antagonized by atropine as well as by chlorisondamine alone. From these results, it is suggested that impulses travelling via the third thoracic ramus or above it mediate myocardial contractile responses in which the nicotinic type response predominates whereaas impulses travelling via the fourth thoracic ramus or below it mediate myocardial contractile responses in which the muscarinic type predominates. additionally, evidence presented indicates that a major proportion of both nicotinic and muscarinic receptors mediating myocardial contractile changes are located in the caudal cervical ganglion rather than the stellate ganglion of the dog.