Changes in CNS myelin proteins and glycoproteins after in situ autolysis. The effects of postmortem autolysis in situ on myelin proteins and glycoproteins were studied in 25- and 125-day-old mouse brain and in adult bovine brainstem. In bovine myelin a loss of the major myelin glycoprotein was the only difference observed when the tissue was left at 19 degrees C for 24 hours compared to immediately frozen material. In the autolysed mouse brain, the myelin major glycoprotein was the most affected component with a 55% decrease. Both myelin basic protein components were degraded with a 35% loss. The other myelin proteins did not change under the conditions used for this study. There was also no change in the specific activity of 2',3'-cyclic nucleotide 3'-phosphohydrolase, a myelin-associated enzyme. Using the double labelling technique with [3H]fucose and [3 5S] sulfate as precursors injected intracranially, a shift of the major myelin glycoprotein labelled with radioactive sulfate towards a smaller apparent molecular size was observed as a result of the autolysis whereas the electrophoretic mobility of the fucose labelled major peak was unaffected.


[Identification by immunofluorescence of various categories of anterior pituitary gland cells (somatotropin, prolactin, corticotropin, melanotropin, gonadotropin, and thyrotropin cells) in the spermophile (Citellus variegatus)]. Using antibodies against human STH, rat prolactin, 1-24, 17-39-synthetic ACTH, alpha- and beta-synthetic MSH, ovine LH, porcine LH beta, bovine TSH saturated with LH, it is possible to identify somatotropic, prolactin, corticotropic, melanotropic, gonadotropic and thyreotropic cells in the adenohypophysis of rodents Citellus variegatus and Graphiurus murinus. The topographical repartition and morphological characters of these cells are described.


Field trial evaluation of a reo-coronavirus calf diarrhea vaccine. Field trials were conducted using an experimental, modified live virus, oral vaccine for prevention of reo- and coronavirus calf diarrhea. Prior to the trials, one or both of the specific causative agents were identified from affected calves in each participating herd. In 21 herds, sequential trials were conducted in which results of uninterrupted vaccination were compared with disease rates during a preceding or subsequent control period. In these herds there was a statistically significant reduction in the morbidity and mortality from disease in 1,598 vaccinates compared with the rates in 829 prevaccination control calves. Morbidity and mortality in 206 post-vaccination control calves rose marginally above the rates in the same vaccinates. In 26 other herds, where double blind trials were conducted, rates of morbidity and mortality from disease were virtually the same for 1,080 vaccinated calves and 355 placebo calves. Vaccinates in the sequential trials had the lowest morbidity and mortality rates of any test group in either field trial format. In a selected dairy herd, both field trial formats were implemented and the results compared. In the double blind trial, vaccinates and placebo calves had comparable rates of morbidity and mortality from disease. When a sequential trial was later implemented, a statistically significant reduction in morbidity and mortality occurred in vaccinates compared with rates in control calves.


An outbreak of infectious laryngotracheitis in commercial poultry flocks in Ontario. An outbreak of infectious laryngotracheitis was observed in Ontario commercial poultry flocks from September 1974 to June 1975. Fifty-five flocks, totalling over 848,770 birds were clinically affected by this disease in the southern region of the province. Sixty-nine percent (38/55) of the exposed flocks were in Wentworth country and the regional municipality of Niagara. Hemorrhagic tracheitis with histological evidence of intranuclear inclusion bodies in syncytia of tracheal epithelial cells were the salient pathological findings.


Neutralizing activity in the gastrointestinal contents of piglets vaccinated with an ethylenimine-inactivated porcine enterovirus. The T80 strain of porcine enterovirus was rapidly and completely inactiviated by ethylenimine in a reaction which appeared to follow first order kinetics. The virus was effectively concentrated 35- to 88-fold, with recovery rates of 23 t0 53%, by adsorption to the polyelectrolyte PE60. Multiple doses of adjuvanted, PE60-concentrated, ethylenimine-inactivated T80 virus given by both the oral and subcutaneous routes induced the appearance of significant levels of virus neutralizing activity in the gastrointestinal tract of piglets vaccinated at four weeks of age. This activity, found predominantly in large intestine, was present 14 days after administration of the first dose of vaccine and significant levels of activity were still detectable six weeks later. Titres of serum virus neutralizing activity were higher and more persistent than in piglets which received live or formaldehyde-inactivated T80 virus by the oral or intramuscular routes.


Demonstration of heavy and light density populations of Aleutian disease virus. A highly purified and concentrated suspension of aleutian disease virus was prepared from large quantities of early infected mink tissues using repeated fluorocarbon extraction procedures. Equilibrium centrifugation of the aleutian disease virus preparation in a cesium chloride gradient yielded three distinct bands at buoyant densities of 1.295, 1.332, and 1.405--1.416 g/cm(3). Electron microscopic observations of these three bands revealed mainly empty particles in the first band. In the second band complete particles with a flattened appearnce predominated and there were also some empty particles. In the third band both complete and empty particles were observed. The size of the aleutian disease virus particles observed in all of the three densities was 23 nm. Light aleutian disease virions (density of 1.332 g/cm3) had a particle to counterimmunoelectrophoresis antigen ratio comparable to that of dense aleutian disease virions (density of 1.405--1.416 g/cm3) but possessed much lower infectivity as determined by mink inoculation.


The significance of cAMP induced alterations in the cellular structure of Phycomyces. Effect of cyclic AMP (cAMP) on Phycomyces blakesleeanus was studied by growing sporangiospores on glucose-asparagine agar or liquid medium containing three different levels of cAMP (10, 20 and 40 micronM) in addition to the control (no cAMP added). The response of Phycomyces to the exogenous cAMP concentration in the medium is as follows: (1) the time required for germ tube emergence is reduced; (2) the diameter of the mycelium is increased (sometimes more than 10 times) and frequency of branching is also increased; (3) the cell wall of the mycelium is thickened (in some cases more than 5 times); (4) the glycogen in the cytoplasm is decreased as visualized in thin sections and also demonstrated in biochemical quantitation; and (5) the distribution of intercalated membranous particles (Imp) on plasma membrane is altered and this can be easily detected in freeze-fractured replica. Such a change in Imp is seen in the formation of small clusters of aggregated particles on the plasmic half (PF) and craters on the complementary exoplasmic half (EF) of the plasma membrane. Although the mechanism of cAMP action requires further exploration, it is possible that the addition of cAMP to the culture medium leads to degradation of glycogen and enhancement of chitin synthesis since the cell wall is largely composed of chitin. The alteration in Imp may be related to a change in the activity of chitin synthetase which is a plasma membrane-bound enzyme.


The effect of N-nitrosobis(2-oxopropyl)amine after oral administration to hamsters. Oral administration of N-nitrosobis(2-oxopropyl)amine (BOP) in drinking water to Syrian golden hamsters for 90 days resulted in a high incidence of intra- and extrahepatic bile duct neoplasms. Only a few pancreatic neoplasms and no lung and kidney tumors developed, in contrast to results obtained after subcutaneous BOP administration.


Presence of ribonucleotide sequences complementary to Rous sarcoma virus (RSV) RNA in chicken cells infected with RSV. RNA--RNA molecular hybridization between [125I] RNA from Rous sarcoma virus virions and RNAs isolated from various subcellular fractions, i.e. nuclei, mitochondria, free and membrane-bound polyribosomes, from tumors induced by RSV in chickens resulted in the formation of RNAase-resistant hybrids only with the RNA of mitochondria and membrane-bound polysomes. The origin of complementary regions in the RNAs from these organelles is discussed.


Hormone binding by human mammary carcinoma. The specific binding of labeled porcine insulin, human prolactin, and human growth hormone was studied in 63 human breast tumors and 15 nonmalignant breast tissues. Most (90%) of the tumors demonstrated significant binding of insulin, as did 80% of nonmalignant tissues. Autoradiographic studies indicated that insulin bound dominantly to tumor cells, rather than to fat and fibrous tissue contained within tumors. Specific binding of prolactin and growth hormone of greater than 1% was seen in 20 and 12% of tumors, respectively, and one tumor studied in detail showed a small amount of saturable, high-affinity prolactin binding. The affinity of binding of insulin and prolactin to tumor was similar to that seen in target tissues in subprimate species (Kd = 4 X 10(-10) M), but the prolactin-binding capacity in the one tumor studied in detail was very low (10 fmoles/mg membrane protein), compared with prolactin-responsive experimental mammary carcinoma.


Levels of mammary tumor virus in hormone-dependent and -independent mouse mammary tumor cells. Levels of mammary tumor virus particles (types A and B) and levels of the virus antigen were assayed in hormone-dependent and -independent mammary tumors of GR mice. Various transplant generations of seven separate tumor lines were investigated. The results indicated that the tumors consisted of different cell clones, each of which exhibited a separate progressive expression and subsequent loss of the mammary tumor virus. When the tumors were transplanted, levels of B particles first declined in the hormone-dependent cells, but in later transplant generations, the B particle content of the autonomous cells also dropped. In some tumor lines, this was accompanied by a decrease in viral antigens and/or A particles, but in other lines these concentrations remained high. One tumor line (line V) that remained hormone-dependent throughout nine transplantations was practically devoid of B particles but contained high levels of A particles and mammary tumor antigen.


Immunoelectron microscopic studies of antibodies in mouse sera directed against mouse mammary tumor virus. Indirect immunoferritin and fixed immunofluorescence tests were carried out on (a) sera of mice hyperimmunized with isologous mouse mammary tumor virus (MMTV) particles or isologous MMTV-producing mammary tumor cells grown in tissue culture and (b) sera of mammary tumor-bearing and tumor-free mice of several inbred strains. Sera were tested against MMTV produced by C3H/HEJ/Tex tissue culture cells (MMT-1). Mammary tumor-bearing A/Dm, C3H/HeTex, and RIII/Dm mice and apparently tumor-free A/Dm mice were found to develop naturally occurring nonprotective anti-MMTV antibodies, whereas sera of tumor-free C3H/HeTex, RIII/Dm, and C57BL/6/Tex female mice and A/Dm, C3H/HeTex, and RIII/Dm male mice did not contain anti-MMTV antibodies. Indirect immunoferritin and fixed immunofluorescence labeling of MMTV particles was prevented by absorbing sera with purified MMTV particles. The results demonstrate the relationship of naturally occurring anti-MMTV antibodies in mouse sera to the presence of mammary tumors, confirm previous reports that mice are not tolerant to MMTV, and further establish the usefulness of the indirect immunoferritin procedure in studies of the immune response of mice.


Alterations in glycosphingolipids of plasma membranes from Morris hepatoma 5123TC. Neutral glycosphingolipids and gangliosides were quantified in lipid extracts from normal rat liver and Morris hepatoma 5123TC and their isolated plasma membranes to determine differences in these components in the cell surface membranes of malignant cells. Glycosphingolipids present in rat liver and hepatoma were concentrated in plasma membranes, and glycosphingolipid patterns in plasma membranes reflected those of their respective whole cells. Neutral glycospingolipids of plasma membranes from normal liver and from hepatoma consisted of ceramide mono- and dihexosides, together accounting for 83 to 86% of the total neutral glycosphingolipids and some ceramide tri- and tetrahexosides. In plasma membranes from hepatoma, the concentration of neutral glycosphingolipids was generally greater than in plasma membranes from normal liver. Gangliosides of plasma membranes from hepatoma were altered more drastically, since trisialogangliosides, present in plasma membranes from normal liver, were absent, while hematosides, monosialogangliosides, and disialogangliosides were increased an average eight-fold. These data are compatible with a concept of incomplete synthesis of trisialogangliosides in hepatoma and an accumulation precursor gangliosides.


Mechanism of action of inosine dialdehyde (NSC 118994) in the inhibition of proliferation of tumor cells in culture. Inosine dialdehyde (INOX), the periodate oxidation product of inosine, inhibited the proliferation of various tumor cell lines in suspension culture in a concentration-dependent manner. A concentration of about 1 mM was required to completely inhibit the proliferation of Novikoff rat hepatoma and mouse L-cells, whereas about 0.1 mM completely inhibited the proliferation of L1210 and P388 mouse leukemia and Chinese hamster ovary cells. INOX inhibited in a similar time- and concentration-dependent manner the synthesis of protein, RNA, and DNA, as measured by the incorporation of labeled amino acid, uridine, and thymidine, into acid-insoluble material, without significantly affecting the incorporation of these precursors into the acid-soluble pool. Flow microfluorometric analyses showed that many of the INOX-treated cells became arrested in G2 + M. The results are consistent with the view that INOX affects multiple metabolic steps. The effects of INOX were quite different from those caused  by typical inhibitors of ribonucleotide reductase, hydroxyurea, and 2,3-dihydro-1H-pyrazolo(2,3-a)imidazole, which very rapidly inhibited DNA synthesis and caused arrest of the cells in G1, with minimal effects on RNA and protein synthesis.


Mutagenicity of cancer chemotherapeutic agents in the Salmonella/microsome test. Seventeen cancer chemotherapeutic agents were tested for their ability to mutate Salmonella typhimurium tester strains in the Salmonella/microsome mutagenicity test. There was a high correlation between the mutagenicity and carcinogenicity of a given agent. Carcinogens positive in the test were Adriamycin, daunomycin, 1-propanol-3,3'-iminodimethanesulfonate, cyclophosphamide, isophosphamide, hycanthone, chlornaphazin, nitrogen mustard, uracil mustard, melphalan, and thio-tepa. Two carcinogesn, actinomycin D and bleomycin, were not detected as mutagens. The presumptive noncarcinogen, methotrexate, was negative in the test. Tilorone and 6-mercaptopurine, tentatively classified as noncarcinogens, were mutagenic. The carcinogenicity of cis-dichlorodiammineplatinum(II), which was positive in the test, has not been determined.


Occurrence of steroid glucuronyltransferases in a hepatoma. Occurrence of estrone, estradiol, and testosterone glucuronyltranferase activities was tested in a well-differentiated hepatoma, Reuber H35. Transferase activities for estrone and estradiol were found in the hepatoma. The Michaelis-Menten kinetics of these two microsomal glucuronyltransferase activities were similar in hepatoma and in liver preparations, except for a somewhat higher apparent Km for estradiol in the hepatoma preparations. Under the same experimental conditions, only trace amounts of testosterone glucuronyltransferase activity could be detected in the hepatoma preparations. By contrast, in liver microsomal preparations, testosterone glucuronyltransferase activity was the highest among the steroid glucuronyltransferase activities tested.


Cell surface markers on epithelial-Burkitt hybrid cells superinfected with Epstein-Barr virus. Attempts were made to superinfect two epithelial-Burkitt hybrid cell lines, designated D98/HR-1 and D98/Raji, with Epstein-Barr virus (EBV) and to investigate the expression of some cell surfacr markers including histocompatibility antigens, and the presence of B-cell markers, such as receptors for the third complement component and for monkey red blood cells. Successful superinfection of D98/HR-1 cells with EBV was made evident by the expression of early antigen and, to a lesser extent, virus capsid antigen. Only a rare D98/Raji cell was found to be positive for early antigen. The histocompatibility antigens of the parental cell lines D98, HR-1, and Raji were expressed on the surfaces of the hybrid cells. Receptors for third complement components b and d were not detected on the hybrid cells or on the D98P OR HR-1 cell lines; they were found, however, on the Raji cells, indicating that EBV receptors and complement receptors can be separated. The significance of the infection of the hybrid cells with EBV and the expression of cell surface markers is described.


[Structure of periodated ribonucleosides and ribonucleotides]. Periodate oxidation converted adenosine, guanosine, cytidine, and uridine into the corresponding dialdehydes. No free dialdehydes are present in solution, but several hydrated species occur (i.r. and n.m.r.). In the solid state, the dialdehydes are completely polymerized. The m.s. molecular peaks corresponding to the free daildehydes were of very low intensity. The c.d spectra of adenosine- and cytidinedialdehyde showed heavily diminished Cotton effects in comparison to the parent nucleosides. For uridine-dialdehyde, the signal intensity of the strongest transition was diminished by about one half, indicating a looser structure allowing free rotation of the base. By contrast, the c.d. signal of guanosine-dialdehyde was increased, indicating self-association of oligomers. The nucleoside-dialdehydes gave, with hydrazides, morpholine derivatives, which posses rigid, stable structures (c.d).


Combined modality treatment for oat cell carcinoma of the lung: a randomized trial 1,2,3. Twenty-three patients with oat cell cancer of the lung were randomized to receive chemotherapy with POCC (procarbazine, 100 mg/m2/day orally, Days 1-14; vincristine, 2 mg iv, Days 1 and 8; cyclophosphamide, 600 mg/m2 iv, Days 1 and 8; and CCNU, 60 mg/m2 orally, Day 1) or COM (cyclophosphamide, 2000 mg/m2 iv, Day 1; vincristine, 2 mg iv, Day 1; and methotrexate, 30 mg/m2 iv, Day 15). After two to three cycles of chemotherapy, all patients were to receive radiotherapy to initially involved sites and then have chemotherapy continued. Patients treated with POCC had a median survival of 14 months vs 10 months for those treated with COM (P = 0.055). Eight of 15 first sites of relapse were intrathoracic and five of these eight had received radiotherapy to that site. Six central nervous system (CNS) failures were evenly divided between the two chemotherapy programs. Thus, the CNS penetration of procarbazine and CCNU in the POCC combination did not prevent CNS relapse. All future patients will receive prophylactic brain radiotherapy.


Structural investigation of Klebsiella serotype K36 polysaccharide. Klebsiella D36 capsular polysaccharide has been investigated by methylation, Smith-periodate oxidation and partial hydrolysis techniques. The structure was found to consist of a hexasaccharide repeating unit as shown. The anomeric configurations of the sugar residues were determined by 1H and 13Cn.m.r.spectroscopy on isolated oligomers obtained during the degradative studies and on the intact polysaccharide.


Chicken macrochromosomes contain an endogenous provirus and microchromosomes contain sequences related to the transforming gene of ASV. Chicken chromosomes from a euploid Marek's lymphoma cell line have been partially fractionated according to size by rate zonal centrifugation in a zonal rotor. DNA-DNA hybridization tests, using unlabeled DNA extracted from gradient fractions and labeled single-stranded, virus-specific DNAs prepared in vitro, indicate that large macrochromosomes harbor the provirus for the endogenous RNA tumor virus of chickens (RAVO), whereas a cellular sequence related to the transforming gene of avian sarcoma virus (ASV) is located in microchromosomes. In support of the method, we have also shown that the single gene for ovalbumin can be assigned to macrochromosomes.


Alkylation of cytosine and 5-hydroxymethyl-cytosine by methyl methanesulphonate and N-methyl-N-nitrosourea: its relevance to mutagenesis. The reaction of cytosine and 5-hydroxymethyl-cytosine (OHMeCyt) with a variety of monofunctional alkylating agents has been investigated to evaluate further the possible role of cytosine alkylation in mutagenesis and the possibility that the immunity of T-even phages to mutation by methyl methanesulphonate (MMS) was due to the unreactivity of OHMeCyt towards this agent. Both cytosine and OHMeCyt reacted equally well with the methylating agents MMS and N-methyl-N-nitrosourea (MNU) affording 6% and less than 1% respectively of the 3-substituted derivative. No product was isolated following subjection of the bases to reaction with ethyl methane-sulphonate (EMS), N-ethyl-N-nitrosourea (ENU) or iso-propyl methane-sulphonate (iPMS).


Comparison of the photochemical and enzymic generation of excited states of oxygen on the induction of benzo(alpha)pyrene mono-oxygenase in liver cell cultures. The photochemical generation of excited states of oxygen such as the superoxide ion(O-2) and singlet oxygen (1o2) by the mild illumination of culture medium containing riboflavin induces benzo(alpha)pyrene mono-oxygenase in 3 different cell lines derived from rat liver. Similar rates of O-2 generation can be produced by the action of xanthine oxidase on xanthine yet this system does not induce the mono-oxygenase. This result confirms that the mono-oxygenase induction is not mediated by O-2 is not mediated by O-2 and that 1O2 is the most likely candidate for stimulating the mono-oxygenase activity.


[Demonstration of specific receptors for angiotensin III in rat adrenal glands]. Distinct and specific binding sites for 3H-angiotensin II (A II) AND 3H-angiotensin III (A III) have been demonstrated in Rat adrenal gland preparations. A III binding sites have the highest affinity (KD29C1-2.10(-10)M) for the 2-8 heptapeptide. This suggests the possibility of a separate and distinct physiological role for the 2-8 heptapeptide, mediated via such A III binding sites.


[Stimulation by ACTH of protein synthesis in cultured adrenal cells. Absence of an effect by cyclin AMP]. ACTH stimulates the incorporation of 14C-leucine into proteins of adrenal tumor cells in culture. This stimulation though about 20% over controls is statistically significant. Cyclic AMP did not reproduce the stimulation observed with ACTH.


Regulation of norepinephrine release from cardiac sympathetic fibers in the dog by presynaptic alpha- and beta-receptors. The effect of phenoxybenzamine (PBA), desmethylimipramine (DMI), clonidine (CLND), sotalol (STL), and isoproterenol (ISPR) on the release of endogenous norepinephrine (NE) from the heart on right cardioaccelerator nerve stimulation was studied in anesthetized dogs. Under control conditions, the catecholamine levels in coronary sinus blood increased linearly with increasing frequencies of stimulation up to 10 Hz and did not increase further at 30 Hz. The release of NE was markedly enhanced after PBA (1 mg/kg, iv) and DMI (1 mg/kg, iv). The enhanced release of NE after DMI, but not after PBA, was associated with a prolonged response in heart rate. In contrast, NE release was reduced after CLND (15 microgram/kg, iv) at stimulation frequencies of 1 and 2 Hz and this was associated with reduced responses in heart rate and left ventricular dtp/dt. STL (5 mg/kg, iv) reduced significantly the release of NE at stimulation frequencies of 1-5 Hz, whereas ISPR enhanced NE outflow at frequencies of 1-4 Hz. These results support the existence of both negative and positive feedback mechanisms on the release of norepinephrine by cardiac sympathetic fibers mediated through presynaptic alpha- and beta-adrenoreceptors, respectively. The functional significance of these mechanisms is also suggested by the correlation found between changes in NE release and variations in cardiac responses under the various drug treatements.


Concentration and composition of the lipoprotein classes in human umbilical cord serum. The concentration and composition of the lipoprotein density classes d less than 1.006 (VLDL), d = 1.006 - 1.064 (LDL), and d = 1.064 - 1.21 (HDL) of human umbilical cord serum were investigated by means of preparative ultracentrifugation. The concentrations of all the density classes, in particular that of the VLDL, are lower than in adults; the values correspond to 8.1 (VLDL), 59.2 (LDL), and 77.4 (HDL) mg/100 ml. About 15% of the total cholesterol and roughly 40% of the total lipid phosphorus did not float at the highest density of 1.21. The composition of the LDL and the HDL was principally in agreement with the pattern obtained in adults. The VLDL of the cord serum, however, showed a lower amount of triglycerides (45%) and a higher content of proteins (22%) than the VLDL from serum of adults. In cord serum only 25% of the total triglycerides are associated with the VLDL fraction, while more than 50% of the triglycerides circulate as a constituent of the LDL.


Plasma triglyceride secretion and metabolism in chronic renal failure. Endogenous very low density lipoprotein triglyceride (VLDL-TG) production rate has been studied in 13 patients with chronic renal failure who were not on dialysis treatment. In these patients the VLDL-TG plasma concentration was significanlty raised when compared with the control group; the fractional turnover rate was reduced but the absolute turnover rate was increased. The results suggest that increased hepatic production of VLDL-TG is a contributory factor to the hypertriglyceridaemia of chronic renal failure.


The combined effect of growth hormone and methandrostenolone on the linear growth of patients with multiple pituitary hormone deficiencies. Six patients with multiple pituitary hormone deficiencies (MPHD) were initially treated with separate courses of methandrostenolone and growth hormone and later with the two drugs combined. During the basal period the mean growth velocity was 2.8 cm/year. Methandrostenolone alone, 0.02-0.05 mg/kg/day given to four of the patients led to an acceleration of the growth velocity to a mean of 5.0 cm/year, while growth hormone 6 mg/week alone accelerated the growth rate to a mean of 6.0 cm/year. Combined therapy led to a striking increase in the mean growth rate to 9.3 cm/year. The shortcoming of the combined growth hormone-androgen therapy was the fast acceleration in skeletal maturation even after short-term administration.


Evaluation of 3 hour metyrapone test in children and adolescents. An evaluation of a new 3 h metyrapone test is presented. The test consists of one oral dose of metyrapone given at 08.00-09.00 hours, and determination of plasma deoxycortisol from a single capillary blood sample taken 3 h later. The test has been assessed in children and adolescents in conjunction with a 5 day metyrapone test, insulin test, vasopressin test, and ACTH test in forty-five reference subjects, in thirty-six hypopituitary subjects with normal or deficient ACTH secretion, in three subjects with primary adrenocortical disease and in ten subjects prior to and after pharmacological prednisone medication. During the first hour after metyrapone the plasma cortisol level decreased to almost the low level maintained for the rest of the 3 h period. The plasma deoxycortisol concentration was basally less than or equal to 35 nmol/l and increased continuously during the 3 h period to the mean level of 299 (95% confidence interval 133-669) nmol/l in the reference subjects. The new test proved to be as accurate as the insulin test in detecting ACTH deficiency. No significant rise was observed in the plasma somatotrophin (GH) level in those children who had a normal GH response to insulin hypoglycaemia.


Human polymorphonuclear leucocytes as mediators of antibody-dependent cellular cytotoxicity to herpes simplex virus-infected cells. Human polymorphonuclear leucocytes (PML) were able to mediate antibody-dependent cellular cytotoxicity (ADCC) against target cells acutely infected with type 1 Herpes simplex virus. The reaction mediated by PML occurred more slowly and required higher concentrations of immune serum than that mediated by human mononuclear cells (MC). At the same ratio of effector cells to target cells, PML-mediated ADCC was less than MC-mediated ADCC. The observed relationship between the number of effector cells added, and the number of target cells lysed, showed that cytolysis mediated by both PML and MC was consistent with 'one hit' probability predictions. This suggested that target cell death resulted from an interaction with a single effector cell. The calculated frequency of effector cells in PML preparation was similar to that in MC, approximately 3-5%. Preliminary examination of the nature of the effector cells suggested that they did not comprise a morphologically distinct subclass of PML. These experiments demonstrate a possible new role for PML in host defence against viral infections.


Smooth muscle antibody in Burkitt's lymphoma and in nasopharyngeal carcinoma. Smooth muscle antibodies (SMA) with specificity for actin, were found with a higher frequency in sera from Burkitt's lymphoma (BL) and nasopharyngeal carcinoma (NPC) patients than in sera from matched controls. No correlation could be found between SMA and anti-Epstein-Barr virus (EBV) antibody titres. There was no parallelism, in individual sera, between the finding of SMA and the occurrence of cold lymphocytotoxins, aother antibody activity found with an abnormally high frequency among BL and NPC patients. The reason why actin, a weak antigen in experimental animals, may become immunogenic in humans remains unexplained.


Inhibition of the in vitro outgrowth of Epstein-Barr virus-transformed lymphocytes by thymus-dependent lymphocytes from infectious mononucleosis patients. Known numbers of thymus-dependent (T) lymphocytes, obtained by positive selection from the blood of acute infectious mononucleosis (IM) patients and from control donors, were added to target cultures of foetal mononuclear cells within 0-7 days of exposure of the target cells to one of a range of doses of Epstein-Barr (EB) virus. The subsequent outgrowth of virus-transformed foetal cells was markedly inhibited by the presence in the cultures of IM-derived T cells, whilst similar numbers of T cells prepared either from cord blood or from adult donors seronegative for EB virus had little or no inhibitory effect. Target foetal cells treated with papain to remove any viral envelope material remaining on the cell surface after infection, were just as sensitive as untreated cells to the addition of IM-derived T cells. It is concluded that the inhibition cannot be mediated through recognition either of viral envelope structures on the surface of infected cells or of the antigenically related virus-determined membrane antigen, MA, but must depend upon recognition of the lymphocyte-detected membrane antigen, LYDMA. The regularity with which IM-derived T cells block the outgrowth of virus-transformed foetal cells suggests that LYDMA consistently appears on the surface of infected foetal cells before the establishment of transformed foci, but is unlikely to be directly associated with the cells' existing histocompatibility antigens.


Non-stedy-state studies of low-density-liproprotein turnover in familial hypercholesterolaemia. 1. The non-steady-state turnover of low-density lipoprotein (LDL), labelled in its apoprotein moiety (apo-B) with 131I, was determined in four patients with familial hypercholesterolaemia, three of them homozygotes. 2. The fractional and absolute catabolic rates (FCR and ACR) of LDL-apo-B were determined by relating the excretion of radioactivity, measured in urine in vitro and by whole-body counter in vivo, to plasma radioactivity and to LDL specific radioactivity respectively. 3. The FCR remained relatively constant, even after marked reduction of LDL pool size by means of plasma exchange. This confirms the existence of an intrinsic defect in LDL catabolism in familial hypercholesterolaemia. 4. LDL-apo-B synthesis, determined by summing the ACR and the daily increment in plasma LDL, was much higher in the three homozygotes than in the one heterozygote, in whom the synthetic rate was normal. 5. These results illustrate the usefulness of combining plasma exchange and whole-body radioactivy counting as a means of examining the relationship between the turnover and pool size of a 131I-labelled protein, such as LDL.


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 by an increase in the number of uterine receptor sites without significant variation in the apparent dissociation constant, which became significant 15 h after nephrectomy. Acute pressor intravenous 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 results provided an explanation for the specific supersensitivity to angiotensin II, observed in uteri excised from nephrectomized rats, which cannot be accounted for by variations in the occupancy of receptor sites. These results also demonstrate that the number of angiotensin receptors, at least in uterine contractile cells, is affected by chronic variations of endogenous angiotensin levels.


Nuclear envelope inclusions demonstrated by freeze-fracture. Inclusions in the perinuclear space of the nuclear envelope of human diploid (MRC-5) fibroblasts, limpet (Patella vulgata) haemocytes, and yeast (Saccharomyces cerevisiae) as observed with the freeze-fracture technique are described. The significance of these inclusions is discussed and it is tentatively concluded that they represent vesicles engaged in transporting macromolecules between nucleus and cytoplasm. Although the inclusions were infrequently observed, their demonstration in mammalian, invertebrate and lower eukaryotic cell types raises the possibility that this form of nucleocytoplasmic exchange may potentially be adopted under appropriate circumstances by the eukaryotic cell in general.


[Criteria for surgical therapy of hypothalamo-hypophyseal Cushing's syndrome]. This article tries to give criteria for the two possible modes of treatment of the pituitary-dependent Cushing's syndrome. Patients (n = 6) who show no secondary complications on Cushing's disease should undergo implantation of 90Y and/or 192Ir into the pituitary gland. After implantation, the excessive secretion of ACTH is suppressed to normal values. The symptoms of Cushing's syndrome disappear. Following the operation, certain patients must also receive hormonal replacement therapy. Fertility is not always preserved. Total adrenalectomy as the alternative procedure is performed in adults with advanced symptoms of the disease (n = 3) and in children (n = 3). Following removal of the adrenal glands, the symptoms of Cushing's syndrome disappear. The appearance of Nelson's tumor after adrenalectomy (10% in the literature) is discussed.


Lacrimal flow.

Lacrimal flow. Syncinetic reflex or isolated orbicularis muscle contraction squeezes the tearsac between Horner's muscle and the inner palpebral ligament, closes the canaliculi and prevents conjunctival fornix backflow, while the tears are expressed (pseudo-peristaltism) towards the nose through the flexible naso-lacrimal canal. The diameter at the level of the canaliculi should be kept optimal in order to allow proper suction towards the tearsac and free capillarity since surface tension varies to a lesser extent. Scopy of contrast lacrimal outflow-channels provides important information on the morphology and the physiopathology of the lacrimal ducts, ruling out eventual functional origin of the lacrimation. The technique is especially important in decision making for treatment (allergic conditions, congenital dacryocystitis) for evaluation of dacryocystorhinostomy, for the early diagnosis and management of infection, inflammation and posttraumatic conditions of the face and anterior skull.


[Herpes simplex causing a therapy-resistant panaritium (author's transl)]. In an eight-month-old child an inflammation developed in the region of the nail bed of the right middle finger. It showed no improvement after immobilisation, extraction of the nail and antibiotics. After ten months unsuccessful treatment electron microscopy of the inflammatory tissue showed numerous Herpes simplex viruses. These increased rapidly in tissue culture and could be neutralised with antibodies against Herpes simplex virus.


Extragenital mixed heterologous tumor of Müllerian type arising in the cecal peritoneum: report of a case. A mixed heterologous tumor of müllerian type arising in the cecal serosa of an elderly woman is reported. There was no history of endometriosis, and no evidence of the latter was found in the surgical specimen or at autopsy. Evidence of mesothelial origin was found microscopically in the adjacent cecal peritoneum. A review of medical literature on mixed heterologous tumors of müllerian type reveals that heretofore they have originated only in the genital system.


[EEG in neuropediatrics (author's transl)]. After remarks about the history of pediatric electroencephalography some particularly important neurophysiological and especially electroencephalographical studies out-of the last ten years are reviewed: polygraphic studies in healthy and sick newborns, spectralanalytical investigations of the EEG in twins, infants and older children. The review is closed by some critical considerations about possibilities and limitations of the automatic analysis of the childhood EEG.


Inhibition of thyrotropin- and dibutyryl cyclic AMP-induced secretion of thyroxine and triiodothyronine by catecholamines. Thyrotropin (TSH), 1 MU/ml and N6, O2'-dibutyryl adenosine 3',5-cyclic monophosphoric acid (dbcAMP) greatly enhanced the release of thyroxine (T4) and triiodothyronine (T3) from mouse thyroids incubated in vitro. L-Epinephrine (E) and L-norepinephrine (NE) strongly inhibited the TSH and dbcAMP-stimulated release of thyroid hormones; L-isoproterenol (IPNE) exerted a relatively weak inhibition. The inhibition by catecholamines was prevented by the alpha-adrenergic blocker, phentolamine; L-propranolol, a beta-adrenergic blocker, had no effect on the inhibition. The TSH-induced release of thyroid hormones was not affected by adrenergic blockers. Epinephrine did not affect the increase in thyroidal cAMP content induced by TSH. These results indicate that catecholamines act by way of an alpha-adrenergic receptor to suppress TSH-stimulated release of thyroid hormones at a point beyond cAMP formation.


Site of action of growth hormone on adrenocortical steroidogenesis in rats. Studies were carried out to define the mechanism of action of growth hormone on adrenocortical steroidogenesis in hypophysectomized female rats. ACTH administration for 7 days increased corticosterone secretion in vivo and corticosterone production by adrenal tissue in vitro. Adrenal mitochondrial and microsomal cytochrome P-450 concentrations as well as the activities of cytochrome P-450-dependent enzymes (cholesterol sidechain cleavage, 11beta-hydroxylase, 21-hydroxylase) were also increased by ACTH. Administration of bovine growth hormone alone to hypophysectomized rats had no effect on any of the parameters evaluated. However, when given in combination with ACTH, growth hormone synergistically enhanced the effects of ACTH on cholesterol sidechain cleavage activity and corticosterone secretion. The magnitude of the pregnenolone-induced difference spectrum in adrenal mitochondria, indicative of cholesterol binding to cytochrome P-450, was also increased by growth hormone, but neither cytochrome P-450 content nor the activities of other steroidogenic enzymes were affected. The results indicate that growth hormone interacts with ACTH to promote corticosterone secretion by increasing the association of cholesterol with adrenal mitochondrial cytochrome P-450, thereby increasing the activity of cholesterol sidechain cleavage, the rate-limiting step in steroidogenesis.


Thyrotropin binding and long acting thyroid stimulator absorbing activities in subcellular fractions from isolated thyroid cells and thyroid homogenates. The ability of various thyroid subcellular fractions to bind [125I]iodo TSH and to absorb long-acting thyroid stimulator (LATS) and thyroid stimulating immunoglobulins (TSI) activities was examined. Membranes purified from thyroid homogenates or isolated thyroid cells absorbed LATS/TSI activities and specifically bound [125I]iodo TSH. Purified thyroglobulin, nuclei, mitochondria and ribosomes did not bind [125I]iodo TSH nor did they absorb LATS/TSI activities. Cell sap obtained by gentle lysis of isolated thyroid cells failed to absorb LATS/TSI activities and to bind labeled hormone. However, freeze-thawing of the cells fragmented the membranes, releasing [125I]iodo TSH binding as well as LATS/TSI absorbing activities into the soluble (cell sap) fraction. The results suggest that the LATS/TSI antigen is of cell surface origin, includes the TSH receptor or larger membrane fragments containing the receptor, and that its release into the soluble fraction is due to the fragmentation of the thyroid membrane during homogenization and preparative procedures.


Effects of prostaglandin E1 and indomethacin on ACTH, prolactin, GH and LH from rat pituitary in vitro. The effects of prostaglandin E1 (PGE1) and indomethacin (IDM) on the release of several pituitary hormones from the rat pituitary were investigated in vitro. An addition of 2 microng/ml of PGE1 to the medium elicited the release of growth hormone (GH) and prolactin, but not of adrenocorticotropin (ACTH) and luteinizing hormone (LH). Although the addition of 1 microng/ml of IDM alone resulted in no effect on the basal release of these hormones, IDM diminished the release of ACTH induced by crude rat hypothalamic extracts (HE) or lysine-8-vasopressin (LVP), and LH induced by HE or luteinizing hormone-releasing hormone (LH-RH). These findings implicate that a part of PGE1 action might be a direct one on the pituitary gland and PGE1 might release GH and prolactin, whereas IDM might have a direct action on the pituitary gland, and that blunt the release of these pituitary hormones induced by several stimuli.


Diagnosis of insulin secreting tumour by retrograde pancreatography. The case of a 26-year-old male with insulinoma is presented. An insulin secreting tumour was indicated by laboratory data. The tumour was accurately localized by endoscopic retrograde pancreatography, as verified by subsequent surgery.


Multicentric early gastric carcinoma mimicking type I (10 years after B-I-surgery). 10 YEARS AFTER B-I-partial gastrectomy for a proven benign gastric ulcer a multicentric early gastric carcinoma type II b, c was detected by endoscopy and histology. This early carcinoma differed from published cases in respect of its macroscopic classification, localization and expansion. The carcinoma situated next to the anastomosis invaded the duodenal mucosa. It also invaded a polypoid fold caused by the previous surgery, thus imitating an early gastric carcinoma type I.


Intrasplenic autotransplantation of canine pancreatic tissues. Maintenance of normoglycaemia after total pancreatectomy. In mongrel dogs, the horizontal part of the pancreas was infiltrated with collagenase, cut in pieces, incubated with collagenase, rinsed twice by centrifugation or sedimentation, and implanted into the spleen of the same animal. The operations were terminated by the removal of the rest of the pancreas. Of 26 operated dogs, one died because of a duodenal perforation, five developed severe hyperglycaemia without remission, and 20 were long-term normoglycaemic survivors followed for up to 10 weeks. These 20 animals became spontaneously normoglycaemic in the course of the first 10 postoperative days. Later, during glucose loading tests, the pattern of blood sugar values was the same in the transplanted animals as in those of a group of non-operated dogs, but the insulin release, although immediate, attained half the control values. The plasma insulin in the splenic vein was more than seven times higher than in the peripheral circulation. Splenectomies performed in seven animals were followed by severe hyperglycaemia and death. Light and electron microscopy demonstrated the presence of the intact endocrine and exocrine pancreatic tissues in the spleens of all animals investigated. It is concluded that laborious separations of endocrine from exocrine tissue are not mandatory for ulterior endocrine function, and that in an animal larger than rodents it is possible to obtain a diabetes-preventing function after the transplantation of only a part of the gland.


Thorotrast-induced renal tumours after retrograde pyelogram. Seven cases of 'thorotrast kidney' after retrograde pyelogram with reflux are dealt with. In five instances cancer of the kidney developed, four of them obvious renal pelvic carcinoma between 14 and 41 years after the thorotrast pyelogram. In some cases the contrast deposits were misinterpreted and in one case the follow-up was also inadequate. There is a high risk of malignancy to connection with 'thorotrast kidneys'. These cases should be follewed up with intravenous pyelogram and exfoliative cytology from renal pelvis. Provided these tests are negative, nephro-ureterectomy should be put off.


Presynaptic alpha-adenoceptors: the depression of self-stimulation by clonidine and its restoration by piperoxane but not by phentolamine or phenoxybenzamine. Depression of self-stimulation by clonidine has been ascribed to continuous direct stimulation of alpha-adrenoceptors with consequent disruption of reinforcement signals thought to be conveyed by noradrenergic pathways. This suggestion was tested by administration of alpha-receptor blocking agents (piperoxane, phentolamine and phenoxybenzamine, PBZ) differing in their affinity for pre- and post-synaptic receptor sites. Piperoxane in low doses (0.55-5.0 mg/kg) previously reported to cause specific blockade of pre-synaptic receptors implicated in negative feedback circuits, caused a significant increase in self-stimulation rate and strongly antagonized the depression of self-stimulation by clonidine (0.15 mg/kg). A larger dose of piperoxane (45 mg/kg) and graded doses of phentolamine and PBZ, affecting both pre- and post-synaptic receptors, depressed self-stimulation, and did not antagonize clonidine-induced depression of self-stimulation. It is concluded that depression of self-stimulation by clonidine may depend on clonidine-induced inhibition of NA release exerted via presynaptic receptors, and that the effect of clonidine is not necessarily evidence that noncontingent adrenergic stimulation disrupts reinforcement.


Discriminative and motor performance in rats after interference with dopamine neurotransmission with spiroperidol. The disruption of instrumental conditioning after block of central dopamine neurotransmission with drugs or lesions has been explained as an inability to initiate movements rather than as a disruption of learning mechanisms per se. To further investigate this we have developed an under-water swim maze which is a test situation that partly counteracts the akinesia of the animals. The maze has been used to test the effect of dopamine receptor inhibition by spiroperidol on a brightness and a spatial discrimination task. Aquisition was blocked and the behaviour randomized after 0.05 mg/kg and higher doses. Swim speed, however, was not affected by 0.05 mg/kg. This suggests that spiroperidol affects the acquisition of this brightness discrimination task independently of its effects on motor performance. Spatial acquisition was not affected even at 5.0 mg/kg which shows that the deficit can not be generalized to all types of acquisition.


Effect of indomethacin on the renal actions of theophylline. The effects of theophylline ethylenediamine (15 mg/kg i.v.) on plasma renin activity (PRA), diuresis, creatinine clearance and plasma and urinary adenosine 3',5'-monophosphate (cyclic AMP) concentrations were studied in urethane-anaesthetized rabbits with and without pretreatment with indomethacin (5 mg/kg i.v.). Theophylline induced a several-fold increase in urinary sodium and water excretion, raised PRA from 76 +/- 15 TO 239 +/- 83 (S.E.M) ng/ml/h (p less than 0.05) and increased urinary cyclic AMP excretion from 0.21 +/- 0.04 to 0.50 +/- 0.08 nmol/min/kg/kidney (p less than 0.05) without any change in arterial plasma cyclic AMP concentration. The ratio between the clearance of creatinine and cyclic AMP was unchanged. After indomethacin pretreatment the theophylline-induced natriuresis was significantly reduced (p less than 0.05), while the effects of theophylline on PRA and urinary cyclic AMP excretion were unchanged. Under the present experimental conditions, indomethacin reduces renal prostaglandin biosynthesis by over 95%. our results indicate that prostaglandins might be concerned with the natriuretic effects of theophylline but not with the effect on PRA.


Serum cholinesterase: function in lipoprotein metabolism. Human serum beta-lipoproteins, isolated by percipitation with heparin-calcium mixture, showed cholinesterase activity. The enzyme activity was almost proportional to the lipoprotein concentration. Rats, treated with neostigmine, a cholinesterase inhibitor, showed a significant decrease in serum beta-lipoprotein and in the incorporation of H3-lysine into the lipoprotein compared to untreated controls. The decreased incorporation of H3-lysine into beta-lipoprotein was associated with increased labelling of alpha-lopoprotein. There was no significant difference in the labelling of pre-beta-lipoprotein. We propose that LDL is formed from VLDL in the presence of cholinesterase.


[Comparative effect of chlorpromazine, imipramine and papaverine on the blood platelet function]. With a view of decoding the mechanisms of their action the effect of papaverine, chlorpromazine and imipramine produced on a number of blood platelets hemostatic functions was studied. All the three drugs suppress in a characteristic fashion the aggregation on blood platelets caused by thrombin in the Tyrode solution and in a plasma defibrilated by heating, as well as by collogen and ADP in the citrated plasma. Unlike chlorpromazine and imipramine papaverine exerts a strong inhibiting action on the phosphodiesterase activity. Chlorpromazine and imipramine suppress the absorption of serotonin and retraction more intensively than this is done by papaverine and call forth morphological changes in the blood platelets that proceed parallel with changes in the intensity of the photodiffusion and liberation of endogenous serotonin. It is postulated that chlorpromazine and imipramine manifest their inhibitory effect through nonspecific damage of the blood platelets membranes, whereas papaverine does this through exchange of adenine-nucleotides and, especially, of 3',5'-AMP.


Perifused adipose cells, quantitation and kinetics of lipolysis. The perifused fat cell system is a system with which lipolytic activity can be monitored on a minute-to-minute basis. Thus, the rate at which lipolysis changes following the addition and removal of hormones can be followed. Catecholamines and other lipolytic agents produced a time-dependent increase in lipolysis following addition of agents, and a time-dependent decrease in lipolysis occurred following removal of the agent. ACTH also produced an increase in lipolysis. However, on termination of ACTH infusion, the lipolytic rate did not return to basal level but remained elevated for at least an additional 30 min (persistent phase). The persistent phase could be terminated by removal of Ca2+. Readdition of Ca2+ in the absence of additional ACTH resulted in a rapid increase in glycerol release. No persistant phase occurred following ACTH if the adipocytes were perifused in a Ca2+-free buffer. However, if Ca2+ was added to the system 20 min after termination of ACTH infusion, lipolysis increased to a rate greater than that obtained initially by infusing ACTH in a Ca2+-free buffer. It is concluded that ACTH is bound to some component of the fat cell in a Ca2+ independent, tenacious manner, and the full manifestation of that binding is dependent on the presence of Ca2+.


Regulation of triglyceride metabolism in the isotopically prelabeled perfused heart. Utilization of 14C-prelabeled endogenous triglycerides was studied in isolated perfused working rat hearts. Lipolysis was estimated by the disappearance of 14C-labeled and total triglycerides. Metabolic 14CO2 production was continuously monitored to evaluate triglyceride fatty acid oxidation. Triglyceride utilization was enhanced by an increase in ventricular pressure development as evidenced by a faster rate of triglyceride mobilization and oxidation. Added catecholamines stimulated lipolysis in hearts perfused with glucose-containing buffer but were without effect in the presence of exogenous fatty acids; the latter were shown to be potent and, possibly, direct inhibitors of myocardial lipolysis. Mediation of catecholamine-induced lipolysis by cyclic AMP has not been settled. Dibutyryl cyclic AMP produced only a slight lipolytic effect, although theophylline, a known phosphodiesterase inhibitor, was a potent lipolytic agent. Theophylline may have exerted its lipolytic effect through an alternative mechanism. Hypoxia per se was a strong inhibitor of heart triglyceride utilization. Furthermore, added epinephrine was without effect on triglyceride lipolysis in hypoxic hearts. Thus, cardiac muscle triglyceride utilization is influenced by such factors as mechanical function, exogenous substrates, hormones, and oxygen availability. The mechanisms involved in these areas of regulation need to be resolved.


Integration of lipid utilization with Krebs cycle activity in muscle. This essay illustrates the ways in which beta-oxidation and the citric acid cycle interact. These included: 1) competition for CoASH, 2) competition for NAD+, and 3) competition for FADH2 oxidation. By means of the above, the cell is able to maintain a precise coordination between the activation of fatty acids in the cytosol, beta-oxidation in the mitochondria, and the complete oxidation of acetyl-CoA to CO2 via the citric acid cycle throughout a wide range of energy demands and oxygen availability.


The effect of shearing of XC DNA on its RSV transfecting activity. XC DNA fragments having the average molecular weight of 10(7) daltons were obtained by hydrodynamic shearing followed by sucrose gradient centrifugation or BUdR treatment of XC cells used for DNA preparation. Such DNA molecules, even after long-term storage, were found to be active in transfection.


The study of the rous virus synthesis in mammalian embryonic tissues growing in vitro. The genome of Rous sarcoma virus was found in mouse and hamster embryo cells growing in vitro shortly after the infection and long before cell transformation occurred. The virus genome was not detected in embryo cells when they were treated with actinomycin D or when the temperature of incubation was lowered to +20 degrees C. This suggested that the synthesis of Rous virus genome could have taken place de novo in the cells investigated. Furthermore, the experiments showed that the life cycle of Rous virus in mammalian and avian cells had at least one phase in common, i.e., that sensitive to the action of actinomycin D.


A double-blind comparative study of Otoseptil Ear Drops and Otosporin Ear Drops in otitis externa. A double-blind comparative study of Otoseptil Ear Drops and Otosporin Ear Drops in otitis externa in fifty-eight patients revealed them both to be equally effective in a series with very few fungal infections. In view of the normally high proportion of infections including fungi, there are obvious advantages in using Otoseptil Ear Drops.


Statistical evaluation of polygraphic investigations of pentoxifylline. Changes in the electro-encephalogram, and in the electro-oculogram electromyogram, ECG, blood supply, blood pressure, electrical skin activity and neurological/psychiatric findings, were investigated in 100 patients given single administrations of 200 mg of pentoxifylline (BL 191). It is concluded from the changes in the EEG wave patterns that pentoxifylline produces a beneficial effect on the cerebral processes contributing to bio-electrical brain activity. Pentoxifylline can be classed as a substance with microcirculatory/metabolic effects on the brain, which lead to stimulation of psychomotor behaviour.


Visual stimulation and wake-sleep behavior in human neonates. Sixteen male and 16 female infants were exposed to 1 of 4 visual stimulus conditions during pre- and postprandial wakefulness. Whereas a nonpatterned gray stimulus minimized the amount of alert inactivity and maximized the amount of subsequent rapid eye movement (REM) sleep, black-and-white checkerboard patterned stimuli produced greater amounts of alert inactivity and correspondingly lesser amounts of subsequent REM sleep. Amounts of alert inactivity and of subsequent REM sleep produced by the checkerboard patterns depended upon pattern size, infant gender, and stimulus presentation strategy. Inversely correlated stimulus control of wakefulness and subsequent REM sleep was proposed as the functional basis of the inversely correlated ontogenesis of wakefulness and REM sleep.


Role of DNA repair in natural resistance of rat ascites hepatomas to nitrogen mustard. Mechanism of natural resistance of rat ascites hepatomas to nitrogen mustard was examined with tumors, especially AH-13 and AH-44, sensitive and resistant to the chemical, respectively. There was found little difference in the uptake and binding of the chemical among the two lines. In contrast, DNA repair activity of the resistant line was higher than that of the sensitive line. This result suggested that natural resistance of AH-44 to the drug may be, at least partially, due to its higher activity of repairing DNA damage.


A case of minimal deviation hepatoma in man with elevated liver-type pyruvate kinase isozyme. A surgical specimen of solitary, encapsulated tumor tissue obtained from a 52-year-old male, diagnosed histologically as well-differentiated hepatocellular carcinoma (Grade II, Edmondson and Steiner) with liver cirrhosis, Type A' (and B is some parts), was found to have a supernormal level of pyruvate kinase Type L and subnormal level of Type M2; the activities (units/mg protein) being 1.21 and 0.12 respectively. The resulting isozyme pattern was apparently "superdifferentiated" as compared with those of not only the tumor-bearing, cirrhotic liver (Type L, 0.19; Type M2, 0.67) but also the normal liver (Type L, 0.47+/-0.05; Type M2, 0.18+/-0.02). The electrophoretic and kinetic properties of the type L isozyme were identical with those of the cirrhotic host liver and a non-cirrhotic control liver. Other enzyme levels in the hepatoma tissue were as follows: Glucose-6-phosphatase, norma; fructose-1,6-bisphosphatase, reduced; glucokinase, absent; and hexokinase Types I and III, and glucose-6-phosphate dehydrogenase, slightly increased. The serum alpha-fetoprotein level was 95 ng/ml. The whole enzyme profile is consistent with the minimal deviation hepatomas in rats. The results were compared with those of other human hepatomas, and the mechanisms of disordered regulation in hepatoma gene expression were discussed.


Relationship between transcerebral passage of tumor cells and brain metastasis. Quantitative study was carried out on transcerebral passage of tumor cells using Yoshida sarcoma and 6 strains of rat ascites hepatoma. The counting was done every second for 40 sec. In relation to the results obtained, histological study was made on the cerebral metastasis of these tumor cells. Metastatic foci in the brain parenchyma, except the meninges and choroid plexus, were formed by island-forming strains (AH-130, AH-272, and AH-7971) which showed comparatively low passage rate and not be single-cell strains (Yoshida sarcoma, AH-7974F, AH-66F, and AH-13) which showed comparatively high rates. Accordingly, it is obvious that there is a correlation between transcerebral passage rates and brain metastasis formation. The mechanisms of metastasis formation were discussed with special reference to arrest of tumor cells in blood vessels.


Altered growth behavior of virus-transformed cells after treatment with dextran sulfate. Growth behavior in vitro of 3T3 cells and the cells transformed by polyoma or SV40 virus was studied after their treatment with dextran sulfate. The transformed cells treated with dextran sulfated showed decreased saturation density compared with untreated cells, whereas saturation density of 3T3 cells was hardly affected by treatment with dextran sulfate. The ratio of saturation density of untreated SV3T3 cells to that of the treated cells was 64.1% at 5 microng/ml and was 43.4% at 10 microng/ml of dextran sulfate. In Py3T3 cells the ratio was 45.0% at both concentrations of 10 and 20 microng/ml. Plating efficiency was not affected in 3 cell lines tested after treatment with dextran sulfate. The treated cells showed a tendency to form thin and not piled-up colonies in the central area. The treated SV3T3 and Py3T3 cells were flattened in shape compared with the untreated cells. In agar medium the treated SU 3T3 and Py 3T3 cells showed the tendency to stop growing after forming small colonies, whereas the untreated cells kept growing anf formed large colonies. These results suggested that dextran sulfate altered temporarily the growth of the transformed cells to that of untransformed cells.


Infectious murine type-C viruses released from human cancer cells transplated into nude mice. Type-C virus particles were revealed by electron microscope in 6 of 9 tumours of cultured and biopsied human cancers heterotransplanted into nude mice. Some tumours in nude mice were explanted for in vitro cultivation. The virus particles were also found in the cultures derived from the virus-positive tumors. They were mostly found extracellularly, but the particles in budding process were also encountered frequently. Cytological study and karyotype analysis of the cultured cells proved these virus-releasing cells as of hus of human origin. From the close correlation between the statistical virus counts and the complement fixation titers for murine gs antigen of the tumors and their cultures, these viruses propagated in human cancer cells were confirmed to be infectious viruses of nude mouse origin. The virus replicating in human cancer cells was readily infected in some of innocent human cancer cells by co-cultivation. It is to be emphasized that infection of animal endogenous viruses on heterotransplanted human cancer cells is a bothersome contamination for human cancer research, especially when searching for a human tumor virus candidate.


Liver protocollagen proline hydroxylase in human liver diseases and experimental liver fibrosis. Liver protocallagen proline hydroxylase activity (PPH activity) was determined in patients with various liver diseases, CCl4-induced liver fibrosis rats and cholin deficiency (tcd) fatty liver rats. The following results were obtained: Liver PPH activity in patients with chronic hepatitis was higher than that in patients with acute hepatitis, while the activity in patients with liver cirrhosis was much higher than that in patients with chronic hepatitis. The activity was higher in patients with chronic active hepatitis than in those with chronic inactive hepatitis. Patients with active and progressive liver cirrhosis were found to have an especially high PPH activity, in whom the activity reflected well the degree of liver fibrosis. Even though fibrosis in persistent hepatitis was almost negligible or slight, the degree of liver PPH activity in persistent hepatitis was similar to that in liver cirrhosis. Liver PPH activities in CCl4-induced liver fibrosis rats and CD fatty liver rats elevated proportionally to the lapse of time. Whilst liver PPH activity in rats of CD fatty liver without fibrosis in 23 to 31 weeks after the start of the experiment was slightly lower than that in rats of CD fatty liver with fibrosis. But liver PPH activity of the former was considerably higher than that of control rats.


Mediation of guinea pig gastric secretion in vitro by cyclic nucleotides. The effect of methacholine, theophylline, and cyclic adenyl and guanyl nucleotides on gastric secretion from antral and proximal duodenal mucosa of the guinea pig was studied. Both 2 mM dibutyryl (db) cAMP and 5 mM theophylline produced significant increases in gastrin secretion, 4.3 +/- 0.7 (P less than 0.001) and 9.3  +/- 2.4 pg mg-1 min-1 (P less than 0.005) respectively, above basal gastrin secretion (1.5 +/- 0.4 pg mg-1 min-1). The combined effect of the two agents was additive (14.5 +/- 3.6 pg mg-1 min-1). Db cGMP (2 mM) had no effect on gastrin secretion. Methacholine produced a dose-related increase in gastrin secretion which at maximum equaled the combined effect of theophylline and db cAMP. The results suggest that gastrin secretion is mediated in part by intracellular cAMP but do not exclude a cooperative involvement of cGMP.


Effects of calcitonin and substance P on the transport of Ca, Na and Cl across rat ileum in vitro. Both salmon calcitonin (SCT) and substance P decreased ileal Na absorption, changed Cl transport from net absorption to net secretion and elevated the short circuit current when added in vitro at concentrations of 10 microng per ml to solutions bathing the serosal surface of rat ileum which had been stripped of its serosal muscle coat. The effects of substance P were of greater magnitude but shorter duration than SCT. Both peptides also increased the bidirectional fluxes of Ca but did not alter net Ca movement. The changes in Na and Cl fluxes and short circuit current are identifical to those which occur when cellular levels of cyclic AMP increased. However, incubation of ileal mucosa with SCT or substance P did not cause a detectable change in cellular levels of cyclic AMP or cyclic GMP. Both the mechanism of action and the possible physiological functions of SCT and substance P in the regulation of electrolyte transport require further investigation. The results with SCT appear to confirm prior suggestions that calcitonin may act directly to produce secretory diarrhea under pathophysiological conditions.


A cytogenetic analysis of cyclic nucleotide phosphodiesterase activities in Drosophila. The genome of Drosophila melanogaster has been surveyed for chromosomal regions which exert a dosage effect on the activities of cAMP phosphodiesterase or cGMP phosphodiesterase. Two regions increase cAMP phosphodiesterase activity when present as duplications. A region of the X chromosome increases cAMP phosphodiesterase activity when duplicated and decreases that activity when deficient. This region has been delimited to chromomeres 3D3 and 3D4, with 3D4 being the most probable locus, and may contain a structural gene for cAMP phosphodiesterase. A region on the third chromosome, 90E-91B, increases cAMP phosphodiesterase activity when duplicated but has no effect on the activity when deficient. Two regions increase cGMP phosphodiesterase activity when present as duplications. A region of the X chromosome, 5D-9C, increases cGMP phosphodiesterase activity when duplicated, but smaller duplications covering this region fail to show such an increase, indicating that a single locus is not responsible for the increase observed for the larger duplication. A region of the third chromosome, 88C-91B, also increases cGMP phosphodiesterase activity when duplicated. Smaller duplications covering this region show smaller increases than that observed for the larger duplication, suggesting that at least three loci between 88C and 91B contribute to the observed increase by that region. Deficiencies covering region 88C-91B do not affect cGMP phosphodiesterase activity. No locus for a presumptive structural gene for cGMP phosphodiesterase has been found. Limitations of the use of segmental aneuploidy in locating structural genes for enzymes are discussed.


The consequences of nullosomy for a chromosomal region affecting cyclic AMP phosphodiesterase activity in Drosophila. A study of Drosophila nullosomic for chromomere 3D4 shows that this region of the genome is necessary for male fertility, normal female fertility and normal oogenesis. Males nullosomic for 3D4 lack normal, motile sperm. Females nullosomic for this region exert a maternal influence on their progeny which results in a diversity of imaginal defects. The observation that chromomere 3D4 is the most probable locus for a chromosomal region which affects cAMP phosphodiesterase activity, and which may contain a structural gene for the enzyme, prompts the hypothesis that the diverse physiological effects caused by nullosomy for 3D4 are the result of an aberrant cAMP metabolism.


The effect of environmental factors of the coking plant on the enzymatic activity of rabbit oral mucosa. Histochemical methods were used to investigate the activities of some intracellular enzymes in the oral mucosa of the rabbits which had been kept in selected work-stands of a coking-plant for 3 months. The findings were compared with the results for the enzymatic activity of the oral mucosa of control rabbits. The epithelium of the oral mucosa of the experimental rabbits was found to be proliferated acanthotically; moreover, there occurred some other morphological changes of the mucosa which often resembled precancerous states of leukoplakia type. In comparison with the control group, the activities of the studied enzymes, i.e. reduced NAD dehydrogenase (NADD), glucose-6-phosphate dehydrogenase (G6PD), lactate dehydrogenase (LDS), succinate dehydrogenase (SD), cytochrome oxidase (CO), and phosphorylase a+b in the epithelium and the connective-tissue cells of the studied mucosa of the experimental animals were as a rule markedly lowered, this decline being especially pronounced for the three last-mentioned enzymes. It was only the proliferating stratum basale of the experimental rabbit epithelium that frequently exhibited enhanced activities of NADD and LDS. Besides, the activities of NADD, G6PD and LDS were of a markedly diverse intensity in the cells of the chaotically proliferating stratum spinosum of the experimental rabbit mucosa. The results point to the noxious modifying effect of chemical and physical agents of the investigated environment on the oral mucosa of the animals studied.


[Effects of several catecholamine-related drugs on aggressive behavior, brain noradrenaline content and tyramine uptake by isolated mice]. Such effects were studied in male albino mice maintained under isolation circumstances for 7 weeks in order to induce aggressiveness. L-DOPA (25 mg/kg) was given concomitantly with DDC (75 mg/kg) or reserpine (0.1 mg/kg), or each was administered signly and intraperitoneally to subjects twice weekly. Two peaks on the aggressive degree were observed at the 3rd-4th and 6th weeks, respectively. At the first peak, mice treated with L-DOPA and/or reserpine demonstrated aggressive behavior to a higher degree than control mice but at the second peak, to a lower degree. Mice treated with L-DOPA and DDC showed the highest degree at the second peak. Tyramine uptake in the brain measured at the 6th week was enhanced in mice treated with combinations of L-DOPA with reserpine or DDC. Noradrenaline content in the brain was lowered in mice treated with L-DOPA and/or reserpine, in comparison with each control value; It is thus concluded that catecholamine-related drugs influence the degree of enhancement of aggressiveness in modes which vary depending on the form of action of each drug.


Study on the partial differential REM deprivation (PDRD). With an aim to make observations on the rebound elevation of REM sleep, we have devised the partial differential REM deprivation (PDRD) method whereby the subjects are awakened forciby at a fixed time early in the morning. By this method, REM sleep is deprivated by about a half and stage 2 sleep by about one third of the baseline value; but stage SWS is not deprivated. When the test set of PDRD is performed repeatedly on an identical subject, the REM sleep time during PDRD night and recovery night is almost always constant. Our PDRD, we believe, is an excellent method that relieves the disconfort of the subject and the physical burden of research workers and yet is capable of measuring almost accurately the magnitude of rebound elevation of REM sleep in each subject.


[Clinical and histopathological studies of cases of lafora-like inclusion bodies]. In case 1, 41-year-old male, developed progressive demetia, paretic gait disturbance and pyramidal signs with the duration of three years. The neuropathological study revealed systemic atrophy as type Pick-disease i.e., lobal atrophy in the frontal and the parieto-occipital regions, degenerative changes in the basal ganglia and in the thalamus, nerve cell loss in the substantia nigra and myelin pallor in the pyramidal tract. Lafora-like inclusions were found in the cerebral cortex and in the cochleal nucleus. In case 2, 45-year-old male, showed character change, cerebellar symptomes and mental deteriotation, and ulcers on the oral mucosa during about 15 years long period. Neuropathological examination showed chronic encephalitis in the brain stem, vacuolar change in the neuron in the olivary nucleus and Lafora-like inclusions in the cochlear nucleus. Though neither generalized conversion nor myoclonus were clinicaly observed in these cases, the inclusions showed histochemically strong similarity with that of the Lafora-disease. These Lafora-like inclusions were compared with those in the literatur, which were reported on various disease of CNS. Finally in respect of predilection of the inclusions, it is likely that the inclusions result from same metabolic disturbance in the cochlear neurons in the Lafora-disease as well as in the present cases.


[Blood protein synthesis in isolated liver cells]. Cells obtained by continuous perfusion with collagenase maintained their ultrastructure and their capacity for protein synthesis in contrast to mechanically isolated rat liver cells. Albumin and angiotensinogen (renin substrate) are synthesized and secreted and the synthesis of both proteins is stimulated by addition of hydrocortisone. As cell suspensions allow the simultaneous investigation of several samples under well defined conditions they can serve as an excellent model for the study of regulatory mechanisms of serum protein synthesis.


Phosphofructokinase (isozymes) activation by theophylline or by 3',5' cyclic AMP administration. Phosphofructokinase activity of rat thyroid, kidney and muscle can be enhanced by in vivo administration of theophylline or cyclic AMP. This enhancement occurs in the different tissues to a different extent depending on the tissue-specific isozymes. Thyroid and muscle phosphofructokinase which is mainly A-type, is strongly stimulated after administering theophylline in the drinking water over a 8 days period. The kidney enzyme which encompasses about 50% of each A and B type is activated to a lesser extent. Liver phosphofructokinase which is pure B-type shows very little response if any. Comparable results have been obtained with either the muscle or liver enzyme after two to four hours treatment with a single injection of 10 mg/rat of either cyclic AMP or theophylline. This short-term activation could be eliminated by treating the animals with Actinomycin D, 30 min prior to theophylline or cyclic AMP. The possible mechanisms leading to the enhancement of phosphofructokinase activity are discussed.


Chemical proof of lipid-protein interactions by crosslinking photosensitive lipids to apoproteins. Intermolecular cross-linkage between high-density apolipoprotein A-I and lecithins and sphingomyelins. The molecular interactions and spatial arrangements of phospholipids and apoproteins of human high-density lipoprotein were studied by a chemical approach. Phosphatidylcholines and sphingomyelins substituted with fatty acyl residues of high specific radioactivity and labelled with the photosensitive azido group in specific positions were prepared by chemical synthesis. They were recombined with apolipoprotein A-I of human serum high density lipoprotein. The lipoprotein complexes containing either azido lecithins or azidosphingo-myelins were purified by agarose chromatography from excess lipids. The irradiation was performed under conditions which prohibit the interference with the apoprotein structure as proven by circular dichroism, fluorescence spectroscopy, immunodiffusion test and disc electrophoresis. Non-covalently bound lipid molecules were removed by Sephadex LH 20 chromatography. Mild alkaline treatment liberated radioactive fatty acids which were not directly linked to the polypeptide chain, but rather via neighbouring phospholipid molecules. The lipoprotein appeared as a single radioactive band in dodecylsulfate polyacrylamide gel electrophoresis as seen by radioscanning, which further proved the covalent linkage of the fatty acyl residues to the polypeptide chain. In the immunodiffusion test, there is no difference between covalently crosslinked phospholipid-apoLp A-I complex and the non-photolyticall treated complex. This is the first chemical proof of the spatial relationship of the hydrophobic side chains of the lipid and polypeptide chains in a lipoprotein complex.


Shutoff of histone synthesis by Mengovirus infection of Ehrlich ascites tumor cells. The histone synthesizing capacity of mengovirus-infected Ehrlich ascites tumor cells and of their corresponding postnuclear supernatants was investigated as a funcion of time post-infection. In addition, histone synthesis was compared with the synthesis of other basic host proteins under identical conditions. In the scope of mengovirus infection of Ehrlich ascites tumor cells the less complex fraction comprising basic protein, separated from the acidic proteins by carboxymethyl cellulose chromatography, can be regarded as a representative of total host protein. Histones and the remaining basic host proteins therefore are well suited as easily identifiable indicators of the host protein synthesizing potential of mengovirus-infected Ehrlich ascites tumor cells. The cessation of histone synthesis proceeds faster than the arrest of the synthesis of other basic host protein.


[A rheological study on plasma lipoproteins (author's transl)]. Lipoproteins of the types LpB and LP-X are studied in a microviscometer to measure intrinsic viscosity. Up to a shear rate of 1700s-1 no shear dependence of viscosity is observed. Intrinsic viscosities are 3.5 and 4.0 for LpB, and 8.5 ml/g for LP-X. Density measurements are used to calculate apparent specific volumes. The results are compatible with a spherical model. An upper limit of 0.45 is estimated for the hydration.


Characterization of immunoglobulins eluted from hamster SV40 tumors. In order to determine if progressively growing tumors specifically bind antitumor antibody, which might play a role in blocking tumor specific cell-mediated immunity, low pH elutions were performed on autochthonous and transplantable SV40 tumors growing in situ for various lengths of time. Ouchterlony analysis revealed that IgG2 was present in all tumor eluates, while IgG1 was present only in eluates from tumors that grew in situ for more than sixty days. IgA and IgM were also found in most eluates but their presence did not correlate with duration of in situ growth. While no antibody to cell surface antigens could be detected, the eluates did possess antibody activity directed to the SV40 nuclear T-antigen. The tumor eluates did not block in vitro lymphocyte dependent microcytotoxicity of immune lymphocytes against cultured SV40 tumor cells. The biological role and significance of different classes of immunoglobulins bound to the surface of growing tumor cells is still poorly understood and requires further study.


Variation between strains of hamsters in the lethality of Pichinde virus infections. Infection by Pichinde virus, a member of the arenavirus group, was studied in Golden Syrian hamsters (Mesocricetus auratus) with regard to possible mechanisms of resistance to virus infection in adult hamsters. Two hamster strains were found to differ in their susceptibility to lethal Pichinde virus infection. LVG/Lak randomly bred hamsters were found to be 100% susceptible to low doses of Pichinde virus during the first 6 days of life, but after 8 days of life, mortality was uncommon. Peak virus titers in the serum of animals infected at 3 days of life were 4 logs greater than in animals infected at 12 days. MHA/Lak inbred hamsters, in contrast, were found to be susceptible to lethal virus infection both as newborns and as adults. Peak virus titers of greater than 10(8) plaque-forming units/ml were observed in serum 8 days after infection of adult MHA hamsters as compared with less than 10(3) plaque-forming units/ml in the serum of adult LVG hamsters. Cultured primary kidney cells and peritoneal macrophages from either hamster strain supported Pichinde virus replication equally well in vitro. Antibodies to the complement-fixing antigens and to antigens at the surface of virus-infected cells were produced by both strains of hamsters. Cyclophosphamide immunosuppression rendered adult LVG animals susceptible to lethal infections, and virus grew to high titers in the treated animals. These findings suggest that immunological factors that appear early in life in LVG hamsters and are deficient in MHA hamsters limit Pichinde virus infection. Unlike previously reported arenavirus diseases, the observations suggest that death is produced by a direct viral effect and not through immunopathological mechanisms.


Polymyxins as inhibitors of polyclonal B-cell activators in murine lymphocyte cultures. The polymyxin antibiotics polymyxin B sulfate and colistin methane sulfonate were examined for their ability to inhibit responses to the polyclonal B-cell activators (PBA) bacterial lipopolysaccharide (LPS), dextran sulfate (DS), pneumococcal polysaccharide (SIII), and purified protein derivative of tuberculin (PPD) in spleen cell cultures. Polymyxin concentrations of 1 and 10 microng/ml significantly inhibited both the deoxyribonucleic acid synthetic and polyclonal antibody responses stimulated by LPS, DS, and SIII. At these concentrations of polymyxins, responses to PPD and to the T-cell mitogens concanavalin A and phytohemagglutinin were not affected. Inhibition was not caused by a generalized lymphocyte toxicity. After dialysis of LPS-polymyxin and DS-polymyxin mixtures, the PBA preparations showed decreased mitogenic activity. Thus, the polymyxins probably interacted directly with the LPS and DS molecules. The mitogenic response to DS was more significantly inhibited than the response to a nonsulfated dextran. The cationic property of the polymyxins probably allows attachment to negatively charged groups in the mitogenically relevant parts of some but not all PBA molecules,, this attachment resulting in the loss of PBA activity.


Characterization of antigens from type A and B yeast cells of Histoplasma capsulatum. The antigenic composition of cytoplasmic extract and culture filtrate antigens of type A and B yeast cells of Histoplasma capsulatum grown in a synthetic medium was studied. These preparations from type A and B yeast cells contained varying amounts of protein, carbohydrates, and protein-carbohydrate complexes. The antigenic analysis of these preparations was performed by antigen-antiserum absorption with subsequent immunodiffusion and cross-immunoelectrophoresis with absorption in situ in an intermediate gel. All protein antigens observed in culture filtrate of either type A or B yeast cells were also present in the cytoplasmic extracts of the same type. The cytoplasmic extract of type A and B yeast cells each contained certain characteristic antigens that were not shared by the other type. Rocket immunoelectrophoresis with different molecular weight fractions of the antigenic preparations from both types indicated the polydisperse nature of Histoplasma yeast cell antigens.


Human nasal epithelial cell culture system: evaluation of response to human interferons. This report describes the response of human nasal epithelial cells to exogenously applied human interferon (IF). To measure the response, a system developed to study human nasal epithelial cells in vitro was used. Nasal epithelial cells responded equally to both leukocyte and fibroblast IF, with development of resistance to virus replication that was time and dose dependent. Cells required contact with 1,000 U of IF (400 69/19 reference units) for 4 h to produce consistently a significant reduction in virus yield. This in vitro cell cultured system may be useful in predicting the in vivo response of human IF in respiratory viral infections.


Biochemical and immunological characterization of deoxythymidine kinase of thymidine kinaseless HeLa cells biochemically transformed by herpes simplex virus type. Thymidine kinase (TK) from herpes simplex virus type 1 (HSV-1) biochemically transformed HeLa cells, purified by affinity chromatography, has been characterized with respect to its electrophoretic mobility, molecular weight, activation energy, substrate specificity, and immunological specificity. TK purified from HSV-1-transformed HeLa cells has the same electrophoretic mobility as TK purified from HeLa cells lytically infected with HSV-1. The sedimentation velocity of purified TK from transformed cells was similar to that previously reported for the lytic enzyme, and its molecular weight was estimated to be 70,000. The activation energy of purified transformed-cell TK was 18.3 kcal/mol. Antiserum prepared against purified HSV-1 TK, although it showed some cross-reactivity, preferentially neutralized homologous TK. The transformed-cell TK antiserum also neutralized the deoxycytidine kinase activity of HSV-1-infected cell extracts but had no effect on deoxycytidine kinase activity of HSV-2-infected cell extract. These results further support the notion that TK acquired by HeLa cells transformed by HSV-1 is of viral and not of cellular origin.


Indirect fluorescent-antibody test for human cytomegalovirus infection in the absence of interfering immunoglobulin G receptors. The presence of immunoglobulin G receptors in human fibroblasts infected with human cytomegalovirus (CMV) resulted in a nonspecific cytoplasmic reaction in the indirect fluorescent-antibody test. Both CMV antibody-positive and antibody-negative sera from human or other animal species produced the cytoplasmic reaction. The substitution of a simian CMV strain for the human virus successfully eliminated this cytoplasmic reaction and, thus, allowed for the observation of virus-induced fluorescent intranuclear inclusions. With the latter system, CMV antibody titers in human sera were equivalent to those obtained by using the human virus and, in addition, allowed for the detection of relatively low-titered serum samples in which antibody measurement was difficult when human CMV-infected cells were used in the indirect fluorescent-antibody test.


Antibody-dependent cellular protection against herpes simplex virus dissemination as revealed by viral plauqe and infectivity assays. Mouse nonimmune peripheral blood lymphocytes (PBL) plus antibody to herpes simplex virus inhibited virus dissemination in herpes simplex virus-infected 3T3 cell culture as revealed by development of viral plaque, size of immunofluorescent foci, appearance of polycaryocytes, and viral infectivity appearing in the culture. These nonimmune PBL did not act alone in inibiting virus dissemination, but did act synergistically in combination with antibody. The ratio of PBL to target monolayer cells needed to produce this effect was 20. Splenic lymphocytes had weak activity, whereas thymocytes were without effect, even in the presence of antibody. Neither interferon nor lymphotoxin was detected in this lymphocyte-mediated response. These findings support previous observations, based on cytotoxicity assays, that antibody-dependent cellular immune processes could be important in control of and recovery from herpesvirus infection.


Direct serological assay for the heat-labile enterotoxin of Escherichia coli, using passive immune hemolysis. Sheep erythrocytes sensitized with the heat-labile enterotoxin (LT) of Escherichia coli exhibited passive immune hemolysis (PIH) when exposed to specific antitoxin and complement. Thus, PIH serves as the basis for an in vitro serological assay for LT that is sufficiently specific and sensitive to differentiate  LT-positive and LT-negative E. coli isolates. The PIH assay for E. coli LT has been performed with the standard Microtiter system and also by a tube method employing the spectrophotometric determination of hemoglobin release. The spectrophotometric method enhances the sensitivity, accuracy, and objectivity of the PIH assay. The increased sensitivity of the spectrophotometric method also facilitates the identification of LT-positive cultures employing polymyxin "mini-extracts" of whole overnight (18 h) broth cultures of 2.0% Casamino Acid-0.6% yeast extract-salts medium rather than mini-extracts of cells derived from 3.5-h subcultures. Thus, large numbers of E. coli isolates can be individually tested for LT in less than 24 h after broth inoculation by a rapid in vitro assay which requires anti-LT serum as the only specific reagent.


Regulation of staphylococcal enterotoxin B. The effect of glucose and the glucose analogues 2-deoxyglucose and alpha-methyl-glucoside on the synthesis and regulation of staphylococcal enterotoxin B was examined. The attenuating effect of glucose on staphylococcal enterotoxin B synthesis was observed. However, when this effect was examined with analogues of glucose, contradictory responses were seen. alpha-Methylglucoside had a slight stimulatory effect on enterotoxin production and other extracellular proteins, whereas 2-deoxyglucose markedly inhibited enterotoxin production. beta-hemolysin and staphylococcal nuclease were also inhibited by 2-deoxy glucose but the synthesis of nuclease could be rescued by the addition of glucose to 2-deoxyglucose-containing cultures. Enterotoxin and beta-hemolysin synthesis were not subject to glucose rescue. The cells used in this study were permeable to cyclic 3',5'-adenosine monophosphate, but the addition of this compound did not reverse glucose repression or 2-deoxyglucose inhibition of enterotoxin B synthesis. We conclude from these data that the regulation of enterotoxin is not under catabolite control as previously reported.


Thymidine kinaseless revertants of Ltk- cells transformed by herpes simplex virus type 1 are resistant to retransformation by homologous virus. Mouse L cells lacking thymidine kinase (Ltk-) that had been transformed to the thymidine kinase-positive (tk+) phenotype by herpes simplex virus type 1 (HSV-1) were cultured in medium containing tritiated thymidine. Six clonal lines of cells surviving this treatment were found to have the following properties: (i) the cells were tk- and had spontaneous back-reversion frequencies to the tk+ phenotype of 10(-5) or less, (ii) the cells contained HSV antigens, although in lesser amounts than in the parental transformed cells, and (iii) the cells were retransformable to the tk+ phenotype by HSV-1 at frequencies of about 1 to 13% of the frequency of the primary transformation of LtK- cells. HSV-1 plaqued as efficiently on monolayers of these cells and replicated in them to the same extent as it did in Ltk- cells. These results indicate that HSV-1-transformed L cells surviving selection with tritiated thymidine are unlike the parental Ltk- cells in that they are damaged in such a way that the cells are resistant to retransformation by homologous virus, although they remain fully permissive for virus replication.


Adenovirus of ring-necked pheasants: purification and partial characterization of marble spleen disease virus. A method for purification of marble spleen disease virus (MSDV) from the spleens of infected turkeys and pheasants is described. It combines chloroform or fluorocarbon extraction with subsequent purification by centrifugation on a cushion of cesium chloride (CsCl). Further purification of MSDV was accomplished with a CsCl equilibrium density gradient. A viral buoyant density of approximately 1.32 to 1.33 g/cm3 was determined. Negative-stain electron microscopy revealed that virus isopycnically banded by CsCl gradient had non-enveloped icosahedral capsids composed of 252 capsomeres. The direct colorimetric diphenylamine assay indicated that MSDV has deoxyribonucleic acid as its nucleic acid. The above evidence demonstrates that MSDV is an avian adenovirus, the first recognized in the ring-necked pheasant (Phasianus colchicus L.).


Affinity of Sendai virus for the inner ear of mice. The direct immunofluorescent antibody method revealed that Sendai virus, which was thought to be most prevalent in respiratory diseases in mice, infected the inner ear of mice when the virus was injected intracerebrally. Specific immunofluorescence that indicated the presence of antigens of Sendai virus was observed in both perilymphatic and endolymphatic structures. The routes of virus spread are discussed. This experimental study, using newborn mice and Sendai virus, may be useful for the investigation of the pathogenesis of viral labyrinthitis not only because of the high frequency of inner ear infection, but also because of long-time survivors which facilitate prolonged observation.


Efficacy of herpes simplex virus type 1 immunization in protecting against acute and latent infection by herpes simplex virus type 2 in mice. ICR mice were immunized with herpes simplex virus type 1 (HSV-1) and later challenged with HSV-2 by footpad inoculation. Both immunized animals and age-matched, nonimmunized controls were observed for ascending neurological disease and latent infection of spinal ganglia resulting from the HSV-2 challenge. Control animals had a 78% incidence of acute and latent infection compared with a 1.7% incidence in immunized mice. The data show immunity to HSV-1 is protective against both acute and latent infection by HSV-2.


Effects of Bordetella pertussis on the sensitivity of inbred mice to vasoactive amines. Pretreatment with Bordetella pertussis was determined to increase significantly the hypovolemia induced by intravenous injections of histamine either alone or in mixture with serotonin in a total of 26 different strains of mice. Two factors affecting the mortality rates observed by challenge after B. pertussis treatment were: the sensitivity of the strains to vasoactive amines before B. pertussis treatment, and their resistance to acute hypovolemic shock. Appropriate crosses and backcrosses between resistant and susceptible strains failed to demonstrate a clear pattern of inheritance of the susceptibility to B. pertussis effects.


Histopathological changes in mice treated with extracts of the histamine-sensitizing factor of Bordetella pertussis. The histamine-sensitizing factor (HSF) of Bordetella pertussis was isolated in a highly purified form. In addition to inducing profound sensitization to histamine, it also caused a significant lymphocytosis and produced an enhancement of reaginic antibody production in animals upon immunization with antigen. Biologically active doses of HSF produced significant pathological changes in mice including congestion and edema of the lung and a marked depletion of cells in the thymus, white pulp of the spleen, and lymph nodes. The lymphocytosis and histological changes in the lungs and lymphoid organs were observed in different strains of mice injected with HSF (even those that are resistant to its histamine-sensitizing effects). Heating HSF at 80 degrees C for 30 min, a treatment that destroys histamine sensitizing and lymphocytosis promoting activities, completely abolishes the ability to induce the changes observed in the lungs and lymphoid organs.


Neutralizing EBV-specific IgA in throat washings of nasopharyngeal carcinoma (NPC) patients. Throat washings from 26 nasopharyngeal carcinoma (NPC) patients from Hong Kong and Tunisia were studied for the presence of transforming EBV. Only six (23%) were found positive which led to the hypothesis of a neutralizing factor in such salivas. The search for EBV-specific antibodies showed that NPC saliva contained neutralizing VCA and EA IgA (54 and 27% respectively) and VCA and EA IgG (73 AND 54% respectively). Both transforming and non-transforming throat washings contained virus particles as visualized by electron microscopy, but in non-transforming salivas (containing IgA and IgG) the particles were found to be clumped. Comparative study of throat washings from patients with Burkitt's lymphoma (BL); infectious mononucleosis (IM), immunodeficiencies, other cancers, and healthy subjects showed that IgA were restricted to NPC cases.