Turnover and tissue distribution of 125-I-labeled low density lipoprotein in swine and dogs. Swine plasma low density lipoprotein (LDL) isolated ultracentrifugally (d 1.019-1.063) was labeled with 125-I, dialyzed, and reisolated by centrifugation at d 1.063. Over 96% of the radioactivity was shown to be associated with the apoprotein. After reinjection into the donor animal, disapperance of 125-I was followed for up to 122 hr. At all time intervals examined, over 95% of the total plasma 125-I was recovered in LDL (D 1.006-1.063), i.e., there was apparently no transfer of radioactivity to high density or very low density lipoproteins. The disappearance curve was biexponential, with half-lives of 0.83 plus or minus 0.06 and 22.5 plus or minus 1.7 hr for the first and second phases, respectively (13 studies). The mean calculated fractional catabolic rate was 0.041 plus or minus 0.003 hr-minus 1. Similar results were obtained in three dogs using autologous LDL of density 1.020-1.050; fractional catabolic rates were 0.031, 0.031, and 0.029 hr-minus 1. Tissue distribution of 125-I was determined in swine killed at various time intervals after [125-I]LDL injection with corrections for radioactivity in trapped plasma. Of the tissues examined, the liver showed by far the highest concentration. Total hepatic radioactivity, expressed as a percentage of total plasma radioactivity, was rather constant and independent of the time of killing from 3 to 122 hr (15.8 plus or minus 1.9%). The total extravascular LDL pool calculated from analysis of the plasma disappearance curves was about 20-30% of the size of the plasma LDL pool. These data are consistent with the conclusion that the liver accounts for a very large fraction of the total extravascular LDL pool. These data are consistent with the conclusion that the liver accounts for a very large fraction of the total extravascular LDL pool and that it is infairly rapid equilibrium with the plasma pool. To what extent the liver is involved in irreversible degradation cannot be inferred from these findings.


Influence of the pituitary gland on insulin secretion in the genetically obese (ob/ob) mouse. The influence of the pituitary gland of lean and genetically obese (ob/ob) mice on insulin secretion from microdissected pancreatic islets of lean and ob/ob mice has been studied by perifusing the pituitaries of these animals in series with the isolated islets and measuring insulin secretion at 5-min intervals over a period of 60 min. It has been shown that the pituitary perifusate of both lean and obese mice stimulate insulin secretion from lean mouse islets but not from obese mouse islets. The maximum stimulation occurs in the first 10 min and with the lean mouse pituitaries returns to the basal level in about 20 min, whereas with the obese mouse pituitaries insulin secretion is about double that from the control islets even after 40 min. A concentration of pure porcine ACTH equivalent to about three times the amount released from the pituitary gland under the experimental conditions used, caused only a small stimulation of insulin release. Possible interpretations of these findings and further lines of investigation are discussed.


Concentrations of cyclic AMP in rabbit ovarian tissue during the preovulatory period and pseudopregnancy after induction of ovulation by administration of human chorionic gonadotrophin. Concentrations of cyclic AMP were measured in rabbit ovaries at various times after injection of an ovulatory dose of human chorionic gonadotrophin (HCG). A biphasic increase in cyclic AMP concentration occurred during the preovulatory period, with peaks 30 min and 3-4 h after HCG injection. Concentrations of cyclic AMP had returned to those observed in ovaries of control oestrous animals before the onset of ovulation 10-12 h after administration of HCG, and remained low throughout the period of pseudopregnancy. Concentrations of cyclic AMP in the newly formed and developing corpora lutea were similar to the concentrations observed in the remainder of the tissue during this period. No significant increase in cyclic AMP concentration was observed 7-9 days after initiation of ovulation. Concentrations of ATP were also investigated during the preovulatory period. The dose-response relationship of HCG to cyclic AMP production in oestrous rabbit ovaries was investigated.


The pituitary-adrenal system of the genetically obese (ob/ob) mouse. Adult genetically obese (ob/ob) mice which are characterized by adrenal hypertrophy and increased secretion of corticosteroids have considerably increased levels of ACTH in the pituitary gland. At 5 weeks of age there is no difference in the pituitary ACTH content of lean and obese animals and dietary restriction, sufficient to maintain body weight at normal values, reduces the pituitary ACTH content of adult obese mice from 14 times the level found in lean litter-mate controls to almost normal values. Using an in-vitro perifusion system, the release of ACTH from isolated pituitary glands was studied. Pituitaries from lean and obese mice responded similarly to stimulation with a crude extract of hypothalamic tissue containing coticotrophin releasing factor (CRF). The CRF content of the hypothalamus in both groups appears to be similar. In contrast with the high pituitary content, plasma values for ACTH in unstressed obese mice are not increased. The results are discussed in relation to other evidence for a hypothalamic disorder in ob/ob mice.


In vitro studies on allotype suppression.  III. compounds of antiallyotype serum active in release from allotype suppression. Spleen cells of b4b6 rabbits, shown to be deficient in their ability to produce b4Ig due to prenatal exposure to anti-b4, formed anti-T2 antibodies marked with the b4 determinant in response to solubilized T2 phage (S-T2) only when cultured in the presence of antibodies specific for the nonsuppressed type (b6), thus confirming and extending the previously reported observation of release from b4 suppression in cultured cells of b4-suppressed b4b5 rabbits treated with anti-b5 serum.  Only antiallotype sera made in b4 rabbits were active in reversing b4 suppression.  Anti-b5 or anti-b6 sera from rabbits of allotypes b6 or b5, respectively, when used in concentrations which completely or partially inhibited the formation of anti-T2 antibodies marked with the corresponding nonsuppressed allotype of the spleen donor, proved to be almost completely ineffective in causing release of suppression.  Exceptions were noted when spleen cells of rabbits advanced in spontaneous escape from suppression were tested with such sera.  The addition of normal b4 serum to non-b4 antiallotypic sera rendered them as effective in releasing b4 suppression in vitro as were antisera from b4 rabbits.  Furthermore, the capacity of a b4 antiallotype serum to cause reversal of b4 suppression could be potentiated by the addition of normal b4 serum, indicating that nonantibody b4 Ig is a limiting factor in such a serum.  Thus, the release from allotype suppression observed in cultures of spleen cells from b4-suppressed heterozygous rabbits is dependent upon the presence of two components: antibodies directed against the nonsuppressed allotype of the donor and normal b4Ig.  These findings are interpreted in terms of alternate hypotheses involving (a) a mechanism of b4 derepression and (b) inactivation of a suppressor cell with recognition for a b4-labeled target.


Secretion of a specific collagenase by stimulated macrophages. Thioglycollate-stimulated mouse macrophages release a specific collagenase into their medium during in vitro culture. The macrophage collagenase has been characterized as a typical metal proteinase which catalyzes the cleavage of the native collagen molecule into three and one-quarter fragments. The extracellular accumulation and low activity in cell lysates suggest that collagenase is a secretion product of the stimulated macrophage. Prolonged secretion of the enzyme at a constant rate for more than 7 days in culture and its inhibition by cycloheximide provide evidence for biosynthesis in vitro. In contrast, secretion of collagenase is barely detectable from unstimulated macrophages which can, however, be stimulated to secret the enzyme by ingestion and intralysosomal storage of latex particles or dextran sulfate. Macrophages laden with latex, an undigestable particle, continue to release collagenase for at least 20 days. Several established mouse cell lines have also been examined for their capacity to secrete collagenase. Collagenase is one of a class of inducible neutral proteinases by which the activated macrophage can modify its extracellular environment.


Elastase secretion by stimulated macrophages. Characterization and regulation. Thioglycolate-stimulated mouse peritoneal macrophages secrete a Proteinase which degrades insoluble elastin. There is little elastase activity in cell lysates but the bulk of the enzyme accumulates extracellularly during culture in serum-free medium. The secretion of elastase is sustained for over 12 days in culture and continued secretion of elastase requires protein synthesis. Unstimulated macrophages secrete very little elastase activity but can be triggered to secrete higher levels of this enzyme by phagocytosis and intracellular storage of latex particles. The macrophages elastase is a distinctive proteinase differing from the elastases of pancreas and granulocytes and is distinct from the other secreted proteinases of macrophages, namely, collagenase and plasminogen activator. The macrophages elastase is a serine proteinase and is inhibited by di-isopropyl phosphoro-fluoridate, ovoinhibitor, EDTA, dithiothretiol, and serum. Its activity is little affected by soybean trypsin inhibitor, turkey ovomucoid and chloromethyl ketones derived from tosyl lysine, tosyl phenylalanine, and acetyltetra alanine. Hydrolysis by macrophage elastase of chromogenic ester substrates for pancreatic elastase could not be detected. Elastase secretion by stimulated macrophages exceeds that by primary and established fibroblast cell strains. It is likely that elastase secretion by macrophages plays a major role in the pathogenesis of chronic destructive pulmonary diseases such as emphysema.


New mutant and congenic mouse stocks expressing the murine leukemia virus-associated thymocyte surface antigen GIX. For several reasons the G(IX) antigen (1) has a prominent place in current work on murine leukemia virus (MuLV): In the prototype G(IX+) mouse strain 129, the G(IX) trait is mendelian, and is expressed selectively (though not exclusively) on thymocytes. Thus, expression of this cell surface component is under the control of cellular genes and is subject to the controls governing the differentiation of T lymphocytes (2). Although the 129 mouse produces no demonstrable leukemia virus such as that found in the AKR strain, it was soon realized that G(IX) antigen must in some way be related to MuLV, because productive infection with MuLV is frequently associated with appearance of G(IX) antigen on cells that are genotypically G(IX-), most notably on MuLV-infected rat cells, or cells that belong to other differentiation pathways (1). The basis of this connection between G(IX) and MuLV has recently become clear from the demonstration that G(IX) is one of MuLV envelope. Therefore, our working hypothesis is that the presence of G(IX) is one of the antigens present on gp69/71 (3,4), the major glycoprotein component of the MuLV envelope. Therefore, our working hypothesis is that the presence of G(IX) antigen always denotes the presence of gp69/71 (though not all variants of gp69/71 need necessarily carry G(IX)). Study of the circumstances under which G(IX) is expressed on the cell surface is thus potentially a powerful approach to understanding how the expression of C-type viral genomes is controlled. Such studies are greatly facilitated by the availability of mutant and congenic strains of inbred mice which differ from the nonmutant or partner strains only with respect to one or another manifestation of the viral genome. It is for this reason that we record here (Table I) some details of two G(IX) mutant and two G(IX) congenic stocks derived in our colonies at Memorial Sloan-Kettering Cancer Center (MSKCC). In addition, to these four strains, Table I includes data for the three relevant partner strains, and for strain AKR, for comparison. These eight strains all differ from one another with respect to one or more MuLV-related traits.


Expression of murine leukemia virus envelope glycoprotein gp69/71 on mouse thymocytes.  Evidence for two structural variants distinguished by presence vs. absence of GIX antigen. Thymocytes of several mouse strains were tested for expression of the gp69/71 envelope component of murine leukemia virus by surface iodination, followed by immunoprecipitation and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis.  Theses strains included two congenic lines differing from their partner stocks with respect to expression of GIX antigen demonstrable in the cytoxicity assay.  We conclude that:(a) two structural variants of gp69/71 can be expressed on mouse thymocytes, (b) these are distinguishable by a small difference in mobility in SDS gels, (c) one carries GIX antigen and the other not, (d) they are coded, or their expression is regulated, by different chromosomal loci that are not closely linked, and (e) both can be expressed together on the thymocytes of inbred mice.  In the intact thymocyte plasma membrane, the sites of group-specific antigen shared by the two gp69/71 variants, unlike the GIX type specificity carried by only one of them, are probably inaccessible to antibody.


Fibroblast surface antigen produced but not retained by virus-transformed human cells. Normal human fibroblasts contain a cell type-specific glycoprotein antigen (SF) that is known to be slowly shed into the medium and to be present also in human serum.  Immunofluorescence with anti-SF antibodies showed that SF antigen has a highly nonrandom fibrillar distribution in surface of normal fibroblasts.  Simian virus 40-transformed fibroblasts also produced the SF antigen, as shown by radioimmunoassay or immunodiffusion tests, but it was not retained by the surface of these cells.  This creates a major difference between the surfaces of normal and malignant cells.


The identification and purification of multiple forms of theta-haemolysin (theta-toxin) of Clostridium perfringens type A. The theta-haemolysin of Clostridium perfringens was purified from culture supernatant fluids of type A strains by fractional ammonium sulphate precipitation and isoelectric focusing in narrow pH 5 to 8 gradients. Four components detected on electrofocusing were designated theta-1(pI6-8to6-9),theta-2(pI6-5to6-6),theta-3(pI6-1to6-3) and theta-4(pI5-7to5-9). Specific activities ranged from 0-4 x 10-6 to 1-2 x 10-6 haemolytic units/mg protein and 2950 to 3600 LD-50/mg protein. Each haemolytic component was activated by cysteine hydrochloride, and inactivated by cholesterol, by addition of sheep erythrocyte ghosts and by heating at 60 degrees C for 10 min; mouse erythrocytes were more resistant than sheep erythrocytes to haemolysis. A reaction of identity was obtained between components in gel diffusion. Sodium dodecyl sulphate polyacrylamide discgel electrophoresis gave molecular weights in the range 59,000 to 62,000 for each component. A similar value was obtained for theta-1 on density gradient ultracentrifugation. Although the multiple forms were free of 11 factors present  in culture supernatants, crossed immunoelectrophoresis and disc gel electrophoresis revealed minor contaminants. These studies reveal that theta-haemolysin has physical properties in common with other oxygen-labile haemolysins.


Nature of the determinant responsible for the adhesion of lactobacilli to chicken crop epithelial cells. Using an in vitro method, some factors affecting the attachment of a strain of lactobacillus to chicken crop epithelial cells have been studied. Time of contact beyond 10 min, pH value, age or growth temperature of the bacterial culture, or nature of the energy source in the growth medium had little or no effect on attachment. Heating to 100 degrees C for 10 min, or treatment with EDTA or surface active compounds was also without effect. Treatment with sodium periodate markedly decreased adhesion, proteolytic enzymes had a smaller effect but wheat germ lipase was completely inactive. The pronounced inhibition of adhesion by periodate suggested the invovement of carbohydrate. However, enzymes known to attack carbohydrate substrates were inactive in reducing adhesion. Concanavalin A, which binds specifically to certain sugar residues, reduced attachment. It is suggested that these concanavalin A receptors on the lactobacillus are responsible for its attachment to crop epithelial cells.


Chemical mutagenesis of Fusarium oxysporum f. sp. lycopersici: non-selected changes in pathogenicity of auxotrophic mutants. Single and multiple auxotrophic mutants of the Fusarium oxysporum f. sp.lycopersici strains which cause Fusarium crown-rot and Fusarium wilt of tomato were obtained by chemical mutagenesis with nitrous acid, nitrosoguandidine and ethylmethaneulphonate. The mutagenesis and selection procedures, adapted for use with a plant pathogenic fungus, are described. Changes in pathogenicity were observed when the auxotrophs were compared with the wild type but no correlation was observed between changes in pathogenicity and the particular nutritional requirement.


Fate of parental simian virus 40 DNA in permissive monkey kidney cells. The fate of parental SV40 DNA in monkey kidney cells was investigated by infecting the cells with purified virus labelled with [3-H]-thymidine. 5-Bromodeoxyuridine (200 mug/ml) was added to the cells at 2 h after infection to label virus progeny DNA. At 72 h post-infection the cells were harvested and virus DNA was extracted and fractionated by isopycnic sedimentation in CsCL solution. The following DNAs with characteristic densities were found: light (LL), 1.70 g/ml; hybrid (HL), 1-75 g/ml and heavy DNA (HH), 1-80 g/ml. Of the total cell-associated [3-H]-radioactivity (derived from parental virus), more than 90% was recovered in unreplicated parental DNA(LL DNA), about 2% was recovered in the HL DNA and about 1-7% was associated with the HH DNA. The unreplicated parental DNA was present as uncoated intact DNA complexed with proteins present in the infected cell. The HL DNA contains one light parental and one heavy progeny DNA strand. The nature of the radioactivity present in the HH DNA remains to be determined.


Polyamine metabolism in cells infected with herpes simplex virus. Ifection of LS cells with HSV-1 resulted in an inhibition of spermidine and spermine synthesis from putrescine, possibly through inhibition of host cell protein synthesis. The rate of putrescine uptake increased soon after infection, and later, polyamines were lost from the cells. Inhibition of spermidine and spermine synthesis by methylglyoxal bis (amidinohydrazone) did not affect virus replication.


Are cytotoxicity and interferon inducing activity of poly(I).poly(C) invariably linked in interferon-treated L cells. Interferon-treated L cells exhibit a specific enhanced susceptibility to the cytotoxic and interferon inducing activities of double-stranded RNAs such as poly(1). poly(C). These activities remained closely linked through widely varying assay conditions, involving, for example, different time anddosage schedules of poly(1). poly (C),suggesting that there is at least one common step in the mechanisms leading to interferon formation and toxicity in interferon-primed cells exposed to poly(1).poly(C). However, some procedures such as addition of metabolic inhibitors (actinomycin D, cycloheximide) and repeated administration of poly(1).poly(C) suppressed the interferon inducing capacity of poly(1).poly(C) without a concomitant decrease of toxicity. Other procedures such as brief treatment of the cells with interferon or DEAE-dextran permitted full expression of the interferon inducing activity of poly(1).poly(C) without any sign of toxicity. The latter results suggest that the mechanisms underlying interferon production and toxicity of poly(1).poly(C) in interferon-treated L cells diverge from a certain point onward.


Ultrastructural aspects of BK virus uptake and replication in human fibroblasts. After exposure of human embryonic fibroblasts to BK virus, virus particles adsorbed to the plasma membrane were engulfed by pinocytosis or captured by vesicles, possibly originating from the endoplasmic reticulum, within 2 h after infection. Most of the virus particles were then transported into lysosomes or into the nucleus, while a small amount of virus was found free in the cytoplasm. Virus particles entered the nucleus between 2 and 12 h after infection, were still detectable in the nucleus at 24 h after infectionand becamse morphologically undiscernible at 30 h after infection, suggesting that a nuclear uncoating mechanism was active between 24 and 30 h after infection. Virus progeny started to appear in the nucleus of infected cells at 4 days after infection, but not until 7 to 8 days after infection did the virus escape into the cytoplasm and cell degeneration became evident. The possible explanations for the long replicative cycle of BK virus are discussed.


Iterferon induction in mice by BHK cells persistently infected with HVJ. Intraperitoneal injection into mice of BHK-HVJ cells (BHK cells persistently infected with HVJ) induced considerable amounts of circulating interferon. Sonication of BHK-HVJ cells almost totally abolished their interferon inducing capacities. This result suggests that a certain native organized structure of virus infected cells might be essential for production of interferon in this system, as is suggested in the in vitro sytem.


Transformation by a temperature sensitive mutant of Rous sarcoma virus in the absence of serum. Cultures of chicken embryo fibroblasts infected with the temperature-sensitive transformation mutant of Rous sarcoma virus, tsLA24PR-A, were arrested between mitosis and S phase by exposure to serum-free medium at the non-permissive temperature (41degree C) for 2 days. On shifting to the permissive temperature (35degree C) the cells assumed a transformed morphology and increased uptake of [2minus 3H]-Deoxy-glucose. There was a concomitant increase in acid insoluble [3H]-thymidine. This suggests that the virus transforming function can cause stationary cells to enter their growth cycle. The level of release of infectious virus was shown to decrease on cell cycle arrest in serum-free medium and not to recover on a shift to 35 degrees, when cellular DNA synthesis and transformation was induced. Cultures rendered stationary in medium containing serum depleted of multiplication stimulating factor did not show this reduction in virus production.


Evidence for a host cell surface antigen on the envelope of avian tumour viruses. Avian sarcoma viruses of the A, B, C, D and E subgroups are inactivated about 100-fold by the serum of rabbits immunized against chick embryo (CE) cells, in the presence, but not in the absence, of complement. The inactivation is not due to the action of the antiserum and complement on the CE cell cultures used for virus assay, nor to anti-Forssman antibodies, but it is presumably due to antibodies to some antigen(s) common to the surface of CE cells and to the virus envelope. This host cell surface antigen (HSCA) is also present on the surface of the helper viruses RAV1 and RAV2 of Bryan strain Rous sacroma virus. However, it cannot be said whether it is identical for viruses of all subgroups. A parallel electron microscopical study has revealed a characteristic swelling and loss of opacity to electrons of virus particles treated with the antiserum and complement, which appears to precede virolysis. Avian sarcoma viruses are not inactivated, in the presence or absence of complement, by antiserum to BHK21 hamster cells transformed by RSV and carrying virus-induced surface antigen (VISA). Therefore, the virus particles do not carry any surface antigen common to transformed non-permissive and permissive cells.


Genetic control of resistance of chick embryo cultures to RSV (RAV 50). The genetic control of resistance of chick embryo cultures to RSV (RAV 50) was studied in crosses between the highly inbred Reaseheath lines, I, C and W and in the test-cross between WC(F1) and RPRL line 7. Embryo cultures resistant to RSV(RAV2) were also resistant to RSV(RAV 50). Genetic analysis of the segregation results of resistance and susceptibility in the F2, back-crosses, and test-cross populations suggests that the tvb genes pleiotropically control the resistance of the embryo cultures to RSV(RAV 50).


Location and abundance of poly (A) sequences in Sendai virus messenger RNA molecules. Adenine-rich sequences from 18S Sendai virus messenger RNA species were 99% adenylate, 3'-OH terminal, and were present in at least 50% of the RNA molecules. Intact virus messenger RNA molecules were resistant to exonucleolytic attack by polynucleotide phosphorylase, suggesting that their 3'-termini are masked.


Replication of reticuloendotheliosis viruses in cell culture: chronic infection. After an initial acute infection with cell killing, chicken or duck embryo fibroblasts infected in culture with reticuloendotheliosis viruses set up a chronic infection with no cell killing or morphological transformation. Essentially all of the chronically infected cells produced virus. The virus production was not sensitive to cytosine arabinoside or mitomycin C as was virus production in an acute infection. The chronically infected cells had a strong group-specific resistancto the c.p.e. of superinfecting reticuloendotheliosis viruses. However, they were sensitive to vesicular stomatitis virus and avian leukosis-sarcoma viruses. After double infection, single cells produced reticuloendotheliosis virus and avian leukosis-sarcoma virus.


Protein synthesis in Sendai virus-infected cells. The rate of protein synthesis in chicken embryo cells infected with Sendai virus 18 to 20 h previously was about two times greater than in mock-infected controls. At this time of infection six stable virus-induced proteins, four major structural proteins (P, NH, NP and M) and two non-structural proteins (28K and 61K), were identified by electrophoresis in SDS-polyacrylamide gel of total cell extracts. The structural glycopeptide F was not detected in the infected cell extracts. Pulse-chase experiments showed that PN NP, M and 28K proteins either did not undergo any post-translational processing or the processing occurred very rapidly. By contrast, a glycopeptide NH was apparently derived from one of two unstable precursons, 69K or 63K, which were revealed only after a short pulse. The synthesis of virus-specific proteins appeared to be regulated since its rate varied for individual classes of proteins. In nucleocapsid-particles isolated from infected cells two major structural proteins (P and NP) were found. A minor component with a very large mol. wt. was revealed in these particles as well as in the virus particle.


Early alteration of poliovirus in infected cells and its specific inhibition. HeLa cells infected with radioactive poliovirus type 2 were disrupted with ultrasonic treatment, followed by addition of a non-ionic detergent. Two types of virus particles were found to sediment at 80 to 90% the rate of native virus. The first of these appeared to be a complex of native virus particles and membrane components, since treatment with 0-2% SDS released infectious native particles. The second was non-infectious and its sedimentation rate was not greatly altered by SDS. One hour after infection this non-infectious particle was the major product of cell-mediated eclipse. We have confirmed that 10 to 30 mg-g/ml S-7, a substituted thiopyrimidine, blocks infection of cells by poliovirus in a specific manner. Analysis of cells infected with radioactive poliovirus type 2 in the presence of S-7 showed that the virus particles remained as the complex which can be disrupted with SDS. In addition to blocking cell-mediated eclipse, S-7 stabilizes poliovirus against heat inactivation in vitro at the same concentrations which block infection. This action resembles the effect of 10-2 M-glutathione, which is also known to block cell-mediated eclipse of poliovirus.


Characterization of adenovirus antibodies by single radial diffusion in agarose gels containing immobilized intact virus particles. The interaction between antibodies and surface antigens of intact immobilized adenovirus particles was studied by single radial immunodiffusion tests (SRDT) in agarose gels. Purified virus particles and antisera against virus particles of selected serotypes representing different subgroups and against structural components of certain serotypes were employed. Antibodies against hexons, pentons and fibres all gave visible zones. Antisera against the latter components were more efficient in forming zones and this was though to be due to the occurrence of relatively smaller amounts of vertex capsomers and fibre antigen at the virus particle surface. The capacity of antibodies against vertex capsomers to form zones was demonstrated in tests with virus particles of intermediate types. The SRDT with immobilized virus particles was found to be highly sensitive for the demonstration of virus particle surface antigens shared between serotypes. Results obtained in previous studies using virus particles haemagglutination-inhibition (HI) tests including anti-antiserum were confirmed. Cross-reactions mainly occurred within subgroups and were ascribed to a sharing of vertex capsomere antigens. In preliminary studies a rapid typing was achieved by examination of paired human sera from five cases of adenovirus infections.


A genetic recombination map of foot-and-mouth disease virus. Sixty ts mutants were isolated from the Pacheco strain of type O foot-and-mouth disease virus after treatment with either 5-fluorouracil or hydroxylamine. The conditions affecting recombination and assay of the ts+ recombinants were standardized. Using two ts mutants resistant to guanidine, three-factor crosses, supported by two-factor crosses, located 34 of the mutations in a linear arrangement. The recombination frequencies between certain pairs of mutations were additive. The guanidine character of the two resistant mutants mapped as a single site mutation and was located near the middle of the map.


Chromatographic studies on picornavirus capsid polypeptides. The polypeptides of encephalomyocarditis, Mouse-Elberfeld and type 5 rhinoviruses behave similarly when chromatographed on calcium phosphate (brushite), each being eluted by a linear phosphate buffer gradient containing sodium dodecyl sulphate in three major peaks, CI, C2 and C3. Analysis of the peaks by polyacrylamide gel electrophoresis suggests that the major capsid polypeptides of these three picornaviruses elute in the order: delta (peak CI), gamma with (peak C2) and alpha (peak C3).


Efficacy of exogenous interferon treatment initiated after onset of multiplication of vesicular stomatitis virus in the brains of mice. Initiation of treatment with potent interferon preparations (6-4 x 10-6 units per dose) 4 days after intranasal inoculation of VSV (at a time when virus has already multiplied in the brains of most mice) resulted in a marked increase in mouse survival. In a series of 6 experiments only 11 to 18% of control mice survived whereas 52% of interferon treated mice survival. These results suggest the usefulness of exogenous interferon even when treatment is begun late in the course of an acute virus disease.


The lact of antiviral effect of (polyinosinic acid): (polycytidylic acid) when attached to insoluble supports. A local antiviral effect can be observed when (poly rI)-(poly rC), bound to Visking discs by u.v. irradiation, is incubated with monolayers of human foreskin fibroblast cells. Radioactive labelling of cytosine residues in (poly rI)-(poly rC) with -125I, has provided a much more sensitive method for determining the fate of the insoluble (poly rI)-(poly rC) than has been available hitherto. The antiviral effect is not related to the amount of (poly rI)-(POLY RC) present on the insoluble support but rather to the amount of polynucleotide lost from the support during incubation. Treatment of (poly rI)-(poly rC) which had been bound to cyanogen bromide-activated Sepharose with eigher dilute alkali or pancreatic ribonuclease released virtually all the polynucleotide. A small amount of (poly rI)-(poly rC) is released from the insoluble matrix in the presence of serum-free Minimum Eagle's Medium.


Degeneration of muscle in association with carcinoma of the bronchus. The clinical features and necropsy findings are described in three cases in which severe and rapidly progressive muscle weakness developed in association with carcinoma of the bronchus. In all three cases, muscular weakness was directly responsible for death. Histological and ultramicroscopical examination in all cases showed an unusual type of degeneration of muscle fibres accompanied by degeneration of intramuscular nerve fibres, but without involvement of the central nervous system, or of the peripheral nerve trunks. The findings are compared with those of previously reported cases, and possible mechanisms for the muscle degeneration are discussed.


Intranuclear "paramyxovirus-like" material in multiple sclerosis, adreno-leukodystrophy and Kuf's disease. Detailed comparative ultrastructural examination of multiple sclerosis (MS) plaques, inflammatory CNS lesions from adreno-leukodystrophy(A-LD), tissue from a case of chronic granulomatous meningitis, biopsy samples of necrotic cerebral cortex and CNS tissue from a case of Kuf's disease (adult-type ceroid lipofuscinosis), has revealed that the intranuclear filamentous material previously thought to be related to a viral infection in MS is a non-specific finding. These intranuclear strands were, however, found in greatest frequency in the acute lesions of MS and were absent from chronically demyelinated areas. The macrophages, lymphocytes and fibrocytes containing filamentous material in the nuclei were mainly perivascular. In A-LD, some macrophages in active lesions contained similar nuclei, and in Kuf's disease they were present in some glial cells in the cerebral cortex.


Cerebral sponginess and GM3 gangliosidosis; ultrastructure and probable pathogenesis. Extensive multifocal vacuolation of the cerebral hemispheres, brain stem, cerebellum, optic nerves and spinal cord were demonstrated in a 3 1/2 month-old infant. This co-existed with marked increases in cerebral and hepatic ganglioside GM3 (hematoside), absence of its higher homologues (GM1 and GM2) and absence of tissue N-acetylgalactosaminyl transferase. Ultrastructurally, there are major abnormalities in myelin and astroglia. The absence of identifiable "storage" material is believed to correlate with an enzymatic defect involved in ganglioside anabolism. A familial occurrence of this disorder is strongly suggested by the clinical history.


Gemistocytic astrocytes in gliomas. An autoradiographic study. Tritiated thymidine (3-H-T) was administered intravenously to seven patients with cerebral gliomas. Autoradiographs of biopsy specimens excised within the next four hours and of autopsy specimens from three of these patients obtained three weeks to six months after the single pulse of 3-H-T revealed the following: (a) no gemistocytic astrocytes and only a few giant astrocytes were labeled in biopsy specimens, despite a high overall labeling index of 5-10% (percentage of cells labeled in the total cell population); and (b) scattered foci of labeled genistocytes occurred in autopsy specimens, despite a sharp drop in overall labeling index. In histologic sections of these same specimens, genistocytes and giant cells occurred as the major cell types in an irradiated tumor, in large clusters near foci of degeneration, and as isolated cells in anaplastic foci. These findings suggest that: 1) gemistocytes and giant astrocytes are similar in origin and growth potential regardless of minor variations in morphology; 2) these cells multiply slowly, if at all, and are closely related to regressive changes within the tumor; 3) these cells may reflect profound proliferative activity in adjacent neoplastic cells; and 4) the labeling index and malignant potential of the tumor as a whole depend upon the more rapidly dividing tumor elements. Thus, if genistocytes and giant cells indicate malignancy, they do so secondarily, and the biologically harmless gemistocyte may be the loser in an intense competition for the substrates needed in cell proliferation.


The third circulation revisited.

The third circulation revisited. The author reviews modern information concerning the formation, flow and functions of the cerebrospinal fluid. Particular attention is given to the lymphatic-like features of the third circulation and to its importance as an internal milieu for nervous tissue.


Continuous intraventricular cerebrospinal fluid pressure recording in hydrocephalic children during wakefulness and sleep. Continuous 24-hour recordings of intracranial pressure and electroencephalographic activity were made on five hydrocephalic children in whom, in the resting wakefulness state, the intracranial pressure (ICP) was considered normal. An increase in both the mean ICP and its oscillations related to cardiac systole was recorded during slow-wave sleep. Further episodic increases, up to a factor of 7 compared to wakefulness values, occurred during sleep. In three patients it was possible to correlate such episodic increases to the rapid eye movement phases of sleep. The authors discuss these phenomena and their possible implication in the progression of hydrocephalus.


Radiation of infratentorial and supratentorial brain-stem tumors. A retrospective survey of 41 patients with unbiopsied thalamic, pontine, and medullary tumors treated with radiation therapy demonstrated 30% to 40% 3-year survival, with seven long-term survivors. Eleven of the 12 patients surviving over 3 years have essentially normal neurological function. Of seven patients in whom histological diagnoses were eventually obtained, four proved to have glioblastomas and three low-grade astrocytomas. Autopsy revealed that all glioblastomatous tumors extended beyond the treatment field; this suggested that larger volumes, perhaps the whole brain, should be irradiated. High doses (at least 5000 rads) should be used since the 3-year survival among those receiving such doses was 45%.


Effect of dietary fat and sucrose on the activities of several rat hepatic enzymes and their diurnal response to a meal. Regulation of the cytoplasmic enzymes, pyruvate kinase (PK), glucokinase (GK), phosphoenolpy ruvate carboxykinase (PEPCK), fructose-1,6-diphosphatase (FDP), ATP citrate-lyase (ATP-CL), NAD-malate dehydrogenase (NAD-MD), NADP-malate dehydrogenase (NADP-MD), glutamic-pyruvic transaminase (GPT), glucose-6-phosphate dehydrogenase (G6PD), and 6-phosphogluconate dehydrogenase (6PGD), in rat liver by dietary fat (F diet) and dietary sucrose (S diet) was investigated. Mealfeeding the S diet to adult rats for 5 and 9 months resulted in a diurnal dietary response (i.e., food response) variation of FDP, GK, ATP-CL, 6PGD, and PK, while meal-feeding the S diet to young rats resulted in diurnal dietary response variation of ATP-CL, G6PD, NADP-MD, 6PGD, GPT, and PK. Meal-feeding the fat diet results in essentially no diurnal variation in enzyme activity. The overall effect of meal-feeding, as compared with ad libitum feeding, of the S diet was to increase the levels of G6PD, ATP-CL, and NADP-MD and to decrease the level of PEck in the meal-fed rats. Young rats meal-fed the two diets have higher enzyme activities than meal-fed adult rats for the observed enzymes (except for GPT and NAD-MD). In general, hepatic levels of the enzymes studied are low in the F diet-fed animals and markedly higher for the S diet-fed animals. These results suggest that dietary carbohydrate specifically induces those enzymes involved in carbohydrate metabolism, whereas dietary fat does not affect their levels. On the basis of prior evidence for an early requirement of RNA synthesis for sucrose induction of G6PD, this widespread induction of liver enzymes by carbohydrate must indicate either increased synthesis of ribosomal RNA with later regulation of synthesis specifically of these enzymes or increased synthesis of a rather large group of specific messenger RNAs i.e., coordinate genetic control of a number of these enzyme messenger RNAs.


Ultrastructure of gastointestinal smooth muscle in ducks with a vitamin E-selenium deficiency. The earliest ultrastructural lesions involving smooth muscle of duck duodenum and gizzard produced by a vitamin E-selenium deficiency were a degereration of sarcoplasmic reticulum and mitochondria. Lysosomes were not observed during these earliest alterations of deficient smooth muscle. Lipid droplets and mineral deposits were present in more severely degernerating smooth muscle that was being infiltrated by heterophils. In areas of necrotic smooth muscle, myoblasts were forming while macrophages, fibroblasts, and an occasional syncytial giant cell contained lipid droplets and surrounded coalesced mineral crystals. Endothelial cells of capillaries and stromal connective tissue were less severely affected with lesions developing after the earliest signs of smooth muscle degeneration. Neuroaxonal degeneration of nonmyelinated nerve fibers was not observed until after most of the smooth muscle had undergone either degeneration or necrosis. A possible explanation for the pathogenesis of the smooth muscle necrosis is discussed in light of the ultrastructural findings.


Capsular and vascular invasion: important prognostic factors in Wilms' tumor. The influence of capsular and vascular invasion on prognosis was determined retrospectively in 52 children with Wilms' tumor. Local recurrence was more prevalent (0.26) in patients with rupture or capsular invasion than in any other group. Inadequate postnephrectomy irradiation or infiltration of the liver by nephroblastoma cells appeared to be predisposing factors in the development of these reccurrences. Of the 26 patients with gross or microscopic vascular invasion, 13 developed metastasis. This proportion was significantly higher than in patients without vascular involvement (p less than 0.05). These observations emphasize the importance of microscopic detection of capsular and vascular invasion in staging of Wilm's tumor.


Substituted thiazolidones: selective inhibition of nicotinamide adenine dinucleotide-dependent oxidations and evaluation of their CNS activity. Eight 2-arylimino-3-(3-N-morpholinopropyl) thiazolid-4-ones were synthesized from the corresponding 1-aryl-3-(3-N-morpholinopropyl) thiocarbamides, characterized, and tested for their effects on the cellular respiratory activity of rat brain homogenates. All substituted 4-thiazolidones selectively inhibited nicotinamide adenine dinucleotide (NAD)-dependent oxidations of pyruvate, citrate, DL-isocitrate, alpha-ketoglutarate, malate, beta-hydroxybutyrate, L-glutamate, and NADH, while the NAD-independent oxidation of succinate remained unaltered. All thiazolidones possessed some degree of anticonvulsant activity against pentylenetetrazol-induced convulsions, and the protection afforded by these compounds at a dose of 100 mg/kg ranged from 30 to 80%. The low toxicity possessed by most of these thiazolidones was reflected by their approximate LD-50 values from 300 mg/kg to greater than 1000 mg/kg. In the present study, the anticonvulsant activity possessed by these substituted 4-thiazolidones was unrelated to their ability to inhibit selectively the NAD-dependent oxidations by rat brain homogenates. These thiazolidones exhibited depression of the CNS activity which, in some cases, was associated with the increase in respiration. All thiazolidones potentiated pentobarbital (sodium) sleeping time in mice when administered in a dose of 100 mg/kg.


Effects of verapamil on myocardial contractility, cardiac adenosine 3,'5'-monophosphate and heart phosphorylase. The effects of verapamil on myocardial isometric force on contraction, cardiac adenosine 3,'5'-monophosphate (cyclic AMP) and heart phosphorylase alpha activity were studied in the isolated perfused rat heart. When hearts were perfused with verapamil (5.98 times 10- minus 8 M), force of contraction was reduced approximately 50% within 4 to 5 minutes; at this point, the concentration of cyclic AMP was significantly lower than control but phosphorylase alpha activity was unchanged. In hearts perfused continuously for 60 minutes with verapamil, force of contraction and cyclic AMP levels returned to normal within 20 minutes after administration of verapamil was begun. Isoproterenol (0.355 nmol/min) reversed the depressant effect of verapamil on cardiac contractility and restored heart cyclic AMP levels to normal. Methoxamine (35.5 nmol/min) given to verapamil-depressed hearts, caused contractile force to return to normal, but cardiac cyclic AMP levels remained low. Mephentermine (23.0 nmol/min) had no effect on cardiac contraction, cyclic AMP or phosphorylase alpha activity in hearts depressed by verapamil. It was concluded that with the concentration of verapamil used in these experiments, the drug caused a transient decrease in force of contraction and myocardial cyclic AMP. Both the depression in myocardial contractility and in cardiac cyclic AMP caused by verapamil were reversed promptly by isoproterenol, whereas methoxamine overcame acutely only the negative inotropic effect of verapamil. Mephentermine had no effect on hearts depressed by verapamil.


Dose-response relationships for arginine vasopressin and synthetic analogs on three types of rat blood vessels: possible evidence for regional differences in vasopressin receptor sites within a mammal. These experiments were undertaken to determine the comparative relationships between chemical structure and biologic (contractile) activity of a series of vasopressin hormone analogs on different rat blood vessels (e.g., aorta, mesenteric arteriole and mesenteric venule). The functional contributions to, and interactions with, phenolic hydroxyl and aromatic groups as well as basicity in positions 2, 3 and 8, respectively, to or with hormone-receptor affinity and intrinsic (contractile) activity, were determined by analyzing the dose-response curves of five vasopressin peptides lacking one or more of these functional groups. The findings demonstrate that: 1) the structure-action relationships for vasopressin peptide-induced contractions on rat blood vessels vary with the particular type of macro- or microvessel (i.e., aorta, arteriole and venule); 2) the phenolic and aromatic groups in positions 2 and 3, respectively, are not only important for hormone-receptor affinity but intrinsic activity as well; 3) the potency (EC50) values for arginine vasopressin as well as the potencies and intrinsic activities of synthetic vasopressin analogs varied between the three different types of rat blood vessels examined; and 4) circulating levels (i.e., 10- minus 11- minus 10- minus 12 M) of [8-arginine]-vasopressin are capable of inducing contractile effects on mesenteric arterioles and venules. The quantitative data obtained in this study support the notion that a heterogeneity of the receptor, which subserves contraction, probably exists in blood vessels within and between vascular beds of a single mammalian species.


Muscarinic cholinergic regulation of cyclic guanosine 3,5-monophosphate in autonomic ganglia: possible role in synaptic transmission. The effects of acetylcholine, and of agents which selectively mimic or block its nicotinic and muscarinic actions, have been studied on the levels of guanosine 3,5-monophosphate (cyclic GMP) and adenosine 3,5-monophosphate (cyclic AMP) in slices of bovine superior cervical ganglion. Low doses of either acetylcholine or the muscarinic agonist, bethanechol, caused a substantial increase in the level of cyclic GMP, and slightly increased the level of cyclic AMP; the acetylcholine- and the bethanechol-induced increases in cyclic GMP were blocked by atropine, the muscarinic antagonist, but not by hexamethonium, the nicotinic antagonist. The nicotinic agonist N,N-dimethylphenylpiperazinium did not alter the level of either of the cyclic nucleotides. Dopamine increased the level of cyclic AMP and acetylcholine partially prevented this dopamine-induced increase in cyclic AMP. The data are consistent with a model for the ganglion in which muscarinic cholinergic synaptic transmission is mediated by an increase in the level of cyclic GMP in the postganglionic neurons and dopaminergic transmission by an increase in the level of cyclic AMP in these neurons.


Effects of pharmacologic agents on release of lysosomal enzymes from alveolar mononuclear cells. Several pharmacologic agents were tested for inhibition of phagocyte-induced release of lysosomal enzymes from guinea-pig alveolar mononuclear cells and peritoneal leukocytes. The activities of the lysosomal enzyme beta-glucuronidase and the cytoplasmic enzyme lactate dehydrogenase were determined in the cells and media following a 2-hour incubation period of cells and zymosan particles. Adrenergic and cholinergic agents, the 3,'5'-adenosine and guanosine cyclic nucleotides, and theophylline had no effect on phagocytosis or release of beta-glucuronidase or lactate dehydrogenase from alveolar cells. Cytochalasin B stimulated lysosomal but not cytoplasmic enzyme release from these cells in the absence of zymosan. Colchicine and hydrocortisone inhibited phagocytosis and beta-glucuronidase extrusion, and phenylbutazone inhibited only beta-glucuronidase release. Colchicine and phenylbutazone also inhibited enzyme release stimulated by cytochalasin B indicating that they were affecting postphagocytic events. The cyclic nucleotides and theophylline were without effect on the peritoneal leukocytes; dexamethasone and phenylbutazone inhibited enzyme release with only the former antagonizing phagocytosis.


Factors affecting the time course of decay of end-plate currents: a possible cooperative action of acetylcholine on receptors at the frog neuromuscular junction. 1. End-plate currents have been studied in gylcerol-treated frog sartorius nerve-muscle preparations with the voltage-clamp technique. 2. Adding the anticholinesterase prostigmine (3 muM) to the solution bathing the muscle caused a 2-7 (mean 3-3) times increase in the time constant of decay of end-plate currents. The anticholinesterase edrophonium (15 muM) also prolonged the time course of end-plate currents. 3. Pre-treatment of the preparation with collagenase, which leads to the removal of acetylcholinesterase in the synaptic cleft, prolongs the time course of end-plate currents. 4. Curare (1-2 muM), cobratoxin (0-13 muM), or alpha-bungarotoxin (0-13-0-26 muM) decreased the time constant of decay of end-plate currents in the presence of prostigmine. 5. These observations are consistant with the suggestion that repeated binding of acetylcholine (ACh) molecules to receptors as the ACh escapes from the synaptic cleft can contribute to the prolongation of end-plate currents which occurrs when acetylcholinesterase activity is eliminated. 6. Increasing the amount of transmitter released from the presynaptic nerve terminal leads to a prolongation of end-plate currents in the presence of prostigmine. 7. In the presence of prostigmine, the second of two end-plate currents (interval 2-10 msec) decays more slowly than the first. 8. ACh (1-40 muM) or carbachol (40 muM) applied in the solution bathing the muscle prolongs end-plate currents in the presence of prostigmine. 9. It is suggested on the basis of the observations described in paragraphs 6 to 8 that the time constant of decay of end-plate currents in the presence of prostigmine increases with increasing concentrations of ACh in the synaptic cleft. In the absence of prostigmine, increasing the concentration of ACh in the synaptic cleft did not change the time constant for decay of end-plate currents. 10. We interpret these results to suggest that ACh can have a cooperative action on receptors such that the association of ACh with one receptor (defined as binding a single ACh molecule) favours the binding or retention of ACh at other receptors. This implies that receptors can interact.


Studies on convulsants in the isolated frog spinal cord. II. Effects on root potentials. 1. In the isolated frog spinal cord picrotoxin, bicuculline, and strychnine were evaluated for their effects on synaptically induced root potentials recorded by the sucrose gap technique. 2. Picrotoxin (greater than 10- minus 4 M) completely blocked the dorsal root potential (DRP) elicited by stimulating the ventral root of the same segment (VR-DRP). Although picrotoxin antagonized the DRP elicited by stimulation of either an adjacent dorsal root (DR-DRP) or the lateral column (LC-DRP), a slower component to these  potentials appeared and increased in size as the concentration of picrotoxin was increased. Thus picrotoxin brings out a later, picrotoxin resistant component to the DR-DRP and LC-DRP. 3. Strychnine (10- minus 8-10- minus 5 M) reduced and abolished the VR-DRP without prolongation and progressively increased and prolonged the DR-DRP (and LC-DRP) and the DR-VRP. Strychnine in higher concentrations (greater than 10- minus 4 M) also reduced the amplitude and prolonged the duration of the compound action potential of afferent fibres. 4. These results combined with those presented in the preceding paper (Barker, Nicoll & Padjen, 1975) suggest that (1) a GABA-like transmitter mediates the final step in the DR-DRP and LC-DRP pathways and that (2) either taurine or beta-alanine may mediate the last step in the VR-DRP pathway.


The sensitivity of adrenal responses to synthetic adrenocorticotrophin in the conscious unrestrained calf. 1. Changes in cortisol and corticosterone output and blood flow from the adrenal gland have been determined in the conscious unrestrained calf during I.V. infusions of synthetic adrenocorticotrophin (Synacthen) at 0-5 ng-kg(-1) min- minus 1 (low dose), 5 ng-kg- minus 1 (medium dose), 50 ng-kg- minus 1 min- minus 1 (high dose) and 500 ng-kg- minus 1 min- minus 1 (medium dose), 50 ng-kg- minus 1 min- minus 1 (high dose) and 500 ng-kg- minus 1 min- minus 1(very high dose). 2. Infusions at the low dose produced a rise in adrenal output of both cortisol and corticosterone to maximum values of approximately 100 and 30 ng-kg- minus 1 min- minus 1 respectively. Mean output of both steroids was significantly increased within 5 min, reached a maximum within 10 min and had fallen to resting levels 10 min after the infusion was discontinued. 3. The effects of infusions at both the high and very high doses were closely similar; maximal cortisol outputs were within the range 600-800 ng-kg- minus 1 min- minus 1 and corticosterone 350-500 ng-kg- minus 1 min- minus 1 in both groups. 4. When the infusions were terminated, pronounced differences were observed in the rates at which steroid outputs declined. Basal levels were restored within 10 min following the low dose and within 60 min in medium dose animals, but both cortisol and corticosterone output were still elevated 2 hr after infusion in high dose animals. In calves infused at the very high dose, cortisol output did not fall significantly during 2 hr period. The ratio of cortisol: corticosterone released from the adrenal gland immediately before infusion (3-2 +/- 0-3) approximated to the proportions in which the two steroids were found in the arterial plasma, but fell progressively to a minimum (1-3 +/- 0-1) with increasing doses of Synacthen. Conversely, the ratio of the steroids in the arterial plasma was increased during infusions at the low dose, but not at the higher doses. 6. No significant change in adrenal blood flow occurred during Synacthen infusion in low dose animals despite the increase in steroid output. In medium dose animals blood flow through the gland rose during infusion by approximately 75 per cent while in both high and very high dose animals the flow increased by up to 300 percent. 7. In the three groups in which adrenal hyperaemia occurred, blood flow had fallen to within the resting range 45 min after infusion: in each case this fall was much more rapid than the fall in steroid output. No significant increase in aortic blood pressure or heart rate accompanied infusion of Synacthen, indicating that adrenal hyperaemia was dependent upon vasodilatation with the gland. 8. Administration of cycloheximide (10 mg/kg) by I.V. injection either before or during an infusion of Synacthen, inhibited steroidogenesis without affecting the vasodilator response.


Two populations or granular vesicles in constricted post-ganglionic sympathetic nerves. 1. A comparative study has been made of the effects of different fixatives on the ultrastructural appearance of granular vesicles accumulated against a constriction in cat hypogastric nerves. 2. After fixation in either osmium tetroxide or in glutaraldehyde followed by post-osmication accumulations of large granular vesicles (65-90 nm in diameter) were observed in profiles of swollen axons. 3. Fixation in acrolein and sodium dichromate revealed a second population of small granular vesicles (mostly 30-50 nm in diameter) in addition to the large vesicles seen after the other fixatives. 4. In reserpinized cats the small granular vesicles were absent. The large granular vesicles were much less numerous, irrespective of the fixative used. 5. It is suggested that either the small vesicles occur in normal noradrenergic axons but are only revealed by catecholamine-sensitive fixatives or they occur as a result of constricting the nerve trunk. Evidence for and against these possibilities is discussed.


Preservation of early discharge of muscle afferents in acrylamide neuropathy. 1. Responses to stretch and the 'early discharge' have been studied in acrylamide neuropathy in the expectation that functional block of the nerve terminals would allow these two processes to be dissociated. 2. Of 538 single group I and group II muscle afferents which were nonresponsive to stretch, ninety-nine displayed 'early discharge' which appeared to be identical with that described in normally functioning muscle stretch afferents. 3. Preservation of 'early discharge' in non-responsive fibres indicates these impulses do not arise from excitation of the muscle spindle itself. The evidence indicates that 'early discharge' is usually ephaptically excited by the muscle action potential.


Inhibition of adrenergic neurotransmission in isolated veins of the dog by potassium ions. 1. In the intact organism, an increase in K+ concentration decreases the reactivity of blood vessels to sympathetic stimulation. The present experiments were designed to determine whether or not K+ interferes with adrenergic neurotransmission. 2. Helical strips cut from dogs' saphenous veins were incubated (4 hr) in Krebs-Ringer solution containing [7-3H]norepinephrine (5 times 10(-8) g/ml). The preparations were mounted for superfusion and isometric tension recording; the superfusate was collected for estimation of total radioactivity and for chromatographic separation of 3H-labelled norepinephrine and metabolites. 3. Supramaximal electric stimulation (5 Hz, 9 V, 2 msec) increased the tension and the [3H]norepinephrine efflux. Increasing the K+ concentration from 5-9 to 1, 15, and 20 m-equiv/l. caused a progressive depression of these contractions and diminished the total 3H efflux in proportion to the relaxation; the decrease in 3H efflux reflected a decrease in intact [3H]norepinephrine. The same increase in K+ concentration did not alter basal tension or basal 3H efflux. 4. Addition of tyramine (4 times 10(-6) g/ml. min) to the superfusate augmented both the tension and the efflux, but these actions were not depresesd by increasing the K+ concentration. 5. Cocaine, phentolamine, and phenoxybenzamine did not prevent the depression by K+ of the response to electric stimulation. 6. These experiments show that K+ causes relaxation of venous smooth muscle constricted by sympathetic stimulation and does so by inhibiting the release of norepinephrine from nerve endings. By contrast, K+ does not inhibit norepinephrine release in response to tyramine. 7. During submaximal electric stimulation (5 Hz, 1-8--3 V, 2 msec), increasing the K+ concentration from 5-9 to 10 and 15 m-equiv/l. potentiated the contractions and increased the [3H]norepinephrine efflux; at 20 m-equil/l, K+ caused transient increases in tension and 3H efflux followed by relaxation and decreased norepinephrine release. After addition of cocaine (10(-5) g/ml. min), K+ only caused relaxation and decrease in 3H efflux, showing that, in addition to inhibition of norepinephrine release, K+ also inhibits the reuptake process. 8. In higher concentrations (40 m-equil/l.), K+ caused both a liberation of norepinephrine and a direct activation of the smooth muscle cells.


The production of neuraminidase by food poisoning strains of Clostridium welchii (C. perfringens). The production of neuraminidase by a classical strain of Clostridium welchii (C. perfringens) type A was studied. Good yields were produced in 5% Proteose Peptone-water medium (PPW5); the enzyme was essentially extracellular but some further neuraminidase could be released by ultrasonic disintegration of the cells. This also released N-acyl neuraminic acid-aldolase (NAN-aldolase) and the degree to which this interferes with the assay for neuraminidase was evaluated. Forty-one British reference food-poisoning strains of C. welchii type A were examined for extracellular neuraminidase production in PPW5. Twelve of 17 strains that produce so-called heat-sensitive spores were neuraminidase positive whereas 20 of 24 strains that are non-haemolytic and produce very heat-resistant sporeswere neuraminidase negative. Variation was found in the ability to produce neuraminidase among strains of a single Hobbs' serotype; four Hobbs' type-13 strains produced neuraminidase but a fifth did not. Disruption of the cells of a Hobbs' type-2 strain that did not produce any extracellular neuraminidase released NAN-aldolase but there was no evidence of cell-associated neuraminidase. British food-poisoning strains of C. welchii type A thus include some that are clearly neuraminidase positive and some that still cannot be shown to produce neuraminidase. There is no correlation between lack of neuraminidase production and the ability to cause food poisoning, although the majority of non-haemolytic heat-resistant strains do not produce neuraminidase. It remains possible that neuraminidase may play a part in C. welchii gas gangrene; it is suggested that the ability to define neuraminidase-negative strains may now be of value in investigating this possibility.


Induction of mutation in Staphylococcus aureus by ethylmethane sulphonate. Staphylococcus aureus strains were treated with ethylmethane sulphonate with the object of obtaining mutants that differed from the wild type in one of the properties that might contribute to virulence. Mutants with isolated loss or deficient production of gamma-haemolysin, coagulase, leucocidin and staphylokinase were obtained. Changes in more than one property were often observed. The loss of delta-lysin production was always coupled with an appreciable reduction in P-V leucocidin production. With that exception, systematised combined loss of a restricted number of staphylococcal characteristics was not observed. Mutants that had lost all characteristics of S. aureus were phage-resistant. Isolated loss of staphylokinase production and deficient P-V leucocidin production were found to be due to the independent loss of two prophages.


The membrane action of antidiuretic hormone (ADH) on toad urinary bladder. Radioactive tracer and electrical techniques were used to study the transport of nonelectrolytes and sodium, respectively, across toad urinary bladders in the presence and absence of ADH. The permeability of lipophilic molecules was roughly proportional to bulk phase oil/water partition coefficients both in the presence and absence of hormone; i.e., ADH elicited a general nonselective increase in the permeation of all nine solutes tested. The branched nonelectrolyte, isobutyramide, was less permeable than its straight-chain isomer, n-butyramide, in control tissues. ADH reduced the discrimination between these structural isomers. Hydrophilic solutes permeated more rapidly than expected. In the presence of hormone, there was no change in the permeation of large hydrophilic solutes considered to move via an extracellular pathway, but there was a marked increase in the permeability of water and other small hydrophilic solutes. Collectively, these results suggest that ADH acts to increase the motional freedom or fluidity of lipids in the cell membrane which is considered to be the preferred pathway for the permeation of lipophilic and small hydrophilic molecules. At concentrations of cAMP and ADH which elicit equivalent increments in the shortcircuit current, the effects of these agents on nonelectrolyte transport and membrane electrical conductance are divergent. Such observations suggest that some membrane effects of ADH may not be directly dependent upon cAMP. ADH in the mucosal solution increased the permeability of the toad bladder when the surface charge on the outer surface of the apical membrane was screened with the polyvalent cation, La-3+. These experiments emphasize that interaction of ADH with membranes of toad urinary bladder may account for at least some effects of this hormone.


Assay for Epstein-Barr virus based on stimulation of DNA synthesis in mixed leukocytes from human umbilical cord blood. Relationships between the rate of DNA synthesis in cultured human umbilical cord leukocytes and the multiplicity of added Epstein-Barr virus (EBV) were studied. At low multiplicities of approximately 0.1 transforming units/cell (approximately 10 physical particles/cell), inoculated cultures demonstrated increased rates of DNA synthesis, by comparison to uninoculated cultures, 3 days after inoculation. Stimulation of DNA synthesis was evident of progressively longer intervals after inoculations of 10-fold dilutions of virus. The rate of DNA synthesis, determined by short [-3H]thymidine pulses, reflected as small as twofold changes in multiplicity and thus can serve as a quantitative assay for the virus. Changes in the rate of DNA synthesis were evident before increases in cell number or alteration in morphology. Stimulation of DNA synthesis in umbilical cord leukocytes was inhibited by treatment of EBV with antibody and also in graded fashion, by progressive doses of UV irradiation to the virus. Induction of DNA synthesis by EBV was serum dependent. Estimates of the number of cells transformed were obtained by extrapolation from a standard curve relating known numbers of transformed cells to [-3H]thymidine incorporation and also by cloning cells after exposure to virus. At the low multiplicities of infection used in these experiments approximately 0.04 to 0.002 of the total cellular population was transformed. The high efficiency of cell transformation by EBV by comparison to other DNA tumor viruses is emphasized.


Evidence for the existence of protomers in the assembly of encephalomyocarditis virus. Two capsid precursor subunits, which sediment on glycerol gradients at 13S and 14S, respectively, have been identified in cytoplasmic extracts of encephalomyocarditis virus-infected HeLa cells. The 13S subunit, which was detected after a 10-min pulse label with -3H-labeled amino acids, contained only capsid precursor chain A (mol wt 100,000). When the 10-min pulse label in such cells was chased for 20 min, the A-containing 13S subunit in the cytoplasmic extracts was replaced by a 14S subunit containing equimolar proportions of three chains: epsilon, gamma, and alpha. This (epsilon, gamma, alpha)-containing 14S subunit could be dissociated into 6S subunits with the same polypeptide composition. The sedimentation properties and the polypeptide stoichiometry of these three precursor subunits, when compared with those of the 13S, (beta, gamma, alpha)(5), and 5S, (beta, gamma, alpha), subunits derived by acid dissociation of purified virions, suggest the following structural assignments: 13S, (A)(5); 14S, (epsilon, gamma, alpha)(5), 6S, (epsilon, gamma, alpha). The molecular weights of the individually isolated capsid chains were determined by gel filtration in 6 M guanidine hydrochloride to be: epsilon, 36,000; alpha, 32,000; beta, 29,500; gamma, 26,500; and delta, 7,800. With the exception of the delta-chain, these values are in reasonable agreement with the values previously determined by electrophoresis on sodium dodecyl sulfatepolyacrylamide gels. These data support the hypothesis that picornavirus capsids are assembled from identical protomers according to the following scheme: (A) leads to (A)(5) leads to (epsilon, gamma, alpha)(5) leads to (delta, beta, gamma, alpha)60-n(epsilon, gamma, alpha)n where n is the number of immature protomers per virion.


Structural polypeptides of rabbit, bovine, and human papillomaviruses. The number and apparent molecular weight of the structural polypeptides of Shope rabbit papilloma virus (RPV), bovine papilloma virus (BPV), and human papilloma virus (HPV) were estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Up to 10 polypeptides were detected in highly purified BPV and HPV full particles; a close homology was found between the polypeptide composition of both viruses. Purified RPV virions gave a similar polypeptide pattern. The main components of the three papillomaviruses are the major polypeptide (VP1) with a mol wt of approximately 54,000 and the three smaller polypeptides (VP8, 9, 10) with mol wt of about 16,500, 15,500 and 12,500, respectively. VP8, VP9, and VP10 are never detected in empty capsids. When BPV virions were disrupted with alkaline buffer, the six lower-molecular-weight polypeptides (VP5 to 10) remained associated with viral DNA. This suggests that they are internal components of the virions and that the four higher-molecular-weight polypeptides (VP1 to 4) may represent external components. The polypeptide compositions of BPV and polyoma virus, another papovavirus, have been compared. The number of BPV and polyoma virus components (10 and 6, respectively) and the molecular weight of their major polypeptide (54,000 and 44,500, respectively) are different; however, the three main DNA-associated polypeptides of BPV (VP8, 9, 10) and the three histone-like components of polyoma virus (VP4, 5, 6) were shown to have identical apparent molecular weights. The possibility that some of the minor components of papillomaviruses may be proteolytic degradation products or cell protein contaiminants is discussed.


Post-transcriptional restriction of human adenovirus expression in monkey cells. Infection of the continuous simian cell lines CV-1 and BSC-1 with human adenovirus type 2 (Ad2) is abortive. However, the restriction of Ad2 reproduction in these cells can be overcome by increasing the Ad2 infectious dose or by coinfection with simian virus 40. Vero, another established simian cell line free of detectable endogenous simian virus 40 DNA, is not restricted in its ability to promote Ad2 growth even at low input multiplicities of Ad2 and in the absence of SV40 helper. The amount of structural Ad2 proteins in total cell extracts of enhanced BSC-1 cells is at least two orders of magnitude higher than that of unenhanced cells. In contrast, comparable quantities of Ad2 mRNA specifying these proteins are found in both the enhanced and the unenhanced cell. Both sets of mRNA can be translated in a cell-free system with equal efficiency.


Selective inhibition of herpes simplex virus by 5-amino-2,5-dideoxy-5-iodouridine. 5-Amino-2,5-dideoxy-5-iodouridine, a nel thymidine analogue, is a potent inhibitor of herpes simplex virus type 1 replication. In contrast to most other nucleoside analogues which possess antiviral activity, 5-amino-2,5-dideoxy-5-iodouridine exhibits little, if any, cellular toxicity. Preliminary evidence suggests that 5-amino-2,5-dideoxy-5-iodouridine selectively inhibits viral-specific DNA synthesis.


Temperature-sensitive mutants of simian virus 40 selected by transforming ability. Eight temperature-sensitive mutants of simian virus 40 which transform rat cells at 32.5 C but not at 38.5 C have been isolated. All the mutants were also temperature sensitive for replication in African green monkey kidney cells and five of them were classified into a single complementation group. No mutant incapable of transforming rat cells at either temperature was isolated.


Characterization of T antigen in cells infected with a temperature-sensitive mutant of simian virus 40. T antigen induced in African green monkey kidney cells by a temperature-sensitive mutant of simian virus 40, defective in a function required for cell transformation, was characterized. The number of T antigen-positive cells estimated by an immunofluorescent techniques was almost equal at permissive (32.5 C) and restrictive (38.5 C) temperatures, but was slightly reduced when the infected cells were incubated at a higher temperature (40.5 C). However, a complement fixation test indicated that the amount of T antigen induced by the mutant is not significantly different from that induced by wild-type virus at 40.5 C. These results suggest that the T antigen-inducing ability of the mutant is not defective. Two distinct molecular species of T antigen were induced by the mutant at the permissive temperature, whereas only one form was observed at the restrictive temperature. The larger molecular form (14 to 15S) induced by the mutant at the permissive temperature was more heat labile than that induced by wild-type virus, suggesting that the mutated gene product is a component of the larger molecular form.


Both NS and L proteins are required for in vitro RNA synthesis by vesicular stomatitis virus. Vesicular stomatitis virions, Indiana serotype, were solubilized with high salt solubilizer and separated by ultracentrifugation into a supernatant fraction containing L, G, NS, and M proteins and pellet fraction containing the RNA complexed with N protein. NS protein was purified from the supernatnat fluid by sequential chromatography on phosphocellulose and diethylaminoethyl cellulose columns. The purified NS protein was assayed in a standard transcription system in combination with purified L protein and purified template (pellet fraction) prepared by renografin or CsCl banding. Results of the polymerase assays indicate that both L and NS proteins are required to reconstitute transcription activity with a highly purified template composed of only RNA and N protein. The NS protein polymerase activity is destroyed by trypsin but withstands 90 C temperatures for 10 min. Cytoplasmic NS protein can substitute for virion NS protein in the in vitro transcription assay.


Recombination between endogenous and exogenous RNA tumor virus genes as analyzed by nucleic acid hybridization. Certain chicken cells that do not spontaneously release virus particles have been shown to produce a subgroup E avian RNA tumor virus, Rous-associated virus 60 (RAV-60), after infection with viruses of other subgroups. The nucleic acids of RAV-60 were analyzed for sequence homologies with the viral nucleic acids contained in the uninfected cell and with those of RAV-2, the exogenous virus used for the preparation of this particular RAV-60 isolate. In addition, these nucleic acids were compared with those of RAV-0, an endogenous virus spontaneously released from line 100 chicken cells. RAV-60 appears to be intermediate between RAV-0 and RAV-2 in its genetic composition, based on the pattern of hybridization obtained with the nucleic acids of these viruses and on the melting profiles of the various hybrid combinations. Of the three viruses tested, RAV-0 appears to have the greatest sequence homology with the viral nucleic acids of the uninfected cell. Hybridization between RAV-60 3-H-labeled complementary DNA and either DNA or RNA from the uninfected cell indicates that RAV-60 contains some nucleic acid sequences which are not present in the cell. In addition, some RAV-60 sequences which hybridize with the cell nucleic acid contain significant amounts of mismatching, as indicated by the lower thermal stability of these hybrid duplexes. Hybrid formation between these partially homologous sequences was excluded under stringent annealing conditions. The data indicate that RAV-60 is a recombinant between exogenous and endogenous viral genes.


Nucleic acid homology of murine xenotropic type C viruses. Two major subclasses of xenotropic (X-tropic) murine type C viruses can be distinguished by nucleic acid hybridization. The most frequently encountered subclass (MuLV-X-alpha) includes isolates from BALB/c, C57BL/6J, C58/J, AKR/J, CBA/J, and DBA/2J inbred strains and from the Asian feral mouse subspecies Mus musculus molossinus. The other subclass (MuLV-X-beta) consists of viruses isolated from the NIH Swiss and NZB/BINJ strains. Thus, significant polymorphism exists among the endogenous type C virogenes of a single species, Mus musculus. MuLV-X-alpha genes are found in strains that also have endogenous mouse-tropic viruses (either N-tropic, B-tropic, or both), whereas the MuLV-X-beta subclass is restricted to mouse strains from which mouse-tropic viruses have not yet been isolated. The results are consistent with a model which proposes that mouse-tropic endogenous viruses are derived from the MuLV-X-alpha subclass.


DNA synthesis in polyoma virus infection. III. Mechanism of inhibition of viral DNA replication by cycloheximide. The formation of viral DNA was inhibited in polyoma virus-infected cells in which protein synthesis had been blocked by cycloheximide. The present studies show the following. (i) The pool of replicating viral DNA molecules was reduced in cycloheximide-treated cells by an amount consistent with inhibition of [3-H]thymidine incorporation into viral DNA, whereas the rate of turnover of the replicating population was not affected. (ii) The rate of conversion of replicating molecules into closed-circular DNA was not affected by cycloheximide. (iii) The rate of elongation of nascent viral DNA fragments into strands of unit genome length was unaffected by cycloheximide. It is concluded that viral DNA synthesis is inhibited in the absence of protein synthesis exclusively at the level of initiation of new rounds of genome replication. Replicating molecules already initiated at the time of addition of cycloheximide matured into progeny closed-circular DNA at a normal rate.


Polyadenylic acid on poliovirus RNA. II. poly(A) on intracellular RNAs. The content, size, and mechanism of synthesis of 3'-terminal poly(A) on the various intracellular species of poliovirus RNA have been examined. All viral RNA species bound to poly(U) filters and contained RNase-resistant stretches of poly(A) which could be analyzed by electrophoresis in polyacrylamide gels. At 3 h after infection, the poly(A) on virion RNA, relicative intermediate RNA, polyribosomal RNA, and total cytoplasmic 35S RNA was heterogeneous in size with an average length of 75 nucleotides. By 6 h after infection many of the intracellular RNA's had poly(A) of over 150 nucleotides in length, but the poly(A) in virion RNA did not increase in size suggesting that the amount of poly(A) which can be encapsidated is limited. At all times, the double-stranded poliovirus RNA molecules had poly(A) of 150 to 200 nucleotides. Investigation of the kinetics of poly(A) appearance in the replicative intermediate and in finished 35S molecules indicated that poly(A) is the last portion of the 35S RNA to be synthesized; no nascent poly(A) could be detected in the replicative intermediate. Although this result indicates that poliovirus RNA is synthesized 5' leads to 3' like other RNA's, it also suggests that much of the poly(A) found in the replicative intermediate is an artifact possibly arising from the binding of finished 35S RNA molecules to the replicative intermediate during extraction. The addition of poly(A) to 35S RNA molecules was not sensitive to guanidene.


Polyadenylic acid on poliovirus RNA. III. In vitro addition of polyadenylic acid to poliovirus RNAs. A crude RNA polymerase preparation was made from HeLa cells infected for 3 h with poliovirus. All virus-specific RNA species labeled in vitro (35S RNA, replicative intermediate RNA [RI], and double-stranded RNA [dsRNA]) would bind to poly(U) filters and contained RNase-resistant stretches of poly(A) which could be analyzed by electrophoresis in polyacrylamide gels. After incubation for 45 min with [3-H]ATP in the presence of the other three nucleoside triphosphates, the labeled poly(A) on the RI and dsRNA migrated on gels as relatively homogenous peaks approximately 200 nucleotides in length. In contrast, the poly(A) from the 35S RNA had a heterogeneous size distribution ranging from 50 to 250 nucleotides. In the absence of UTP, CTP, and GTP, the size of the newly labeled poly(A) on the dsRNA and RI RNA was the same as it was in the presence of all four nucleoside triphosphates. However the poly(A) on the 35S RNA lacked the larger sequences seen when the other three nucleoside triphosphates were present. When [3-H]ATP was used as the label in infected and uninfected extracts, heterogeneous single-stranded RNA sedimenting at less than 28S was also labeled. This heterogeneous RNA probably represents HeLa cytoplasmic RNA to which small lengths of poly(A) (approximately 15 nucleotides) had been added. These results indicate that in the in vitro system poly(A) can be added to both newly synthesized and preexisting RNA molecules. Furthermore, an enzyme capable of terminal addition of poly(A) exists in both infected and uninfected extracts.


Presence of herpes simplex virus-related antigens in transformed L cells. Antiserum prepared against herpes simplex virus type 1 (HSV-1)-infected L cells, i.e., lytic antiserum, was shown by an indirect immunofluorescence test to stain 90 percent of HSV-transformed L or HeLa cells. Immunofluorescence in these cells was always most intense in the perinuclear cytoplasmic region. Similar results were obtained with antiserum prepared against HSV-transformed L cells. These data indicate that HSV-transformed cells (both L and HeLa) express HSV-related antigens. Antiserum prepared against HSV-1-transformed L cells, i.e., transformed-cell antiserum, was found to agglutinate purified HSV type 1 virions but failed to neutralize infectivity. This suggests that HSV-1 structural antigens are expressed in HSV-1-transformed L cells. Immunodiffusion studies showed that at least two HSV-related antigens could be demonstrated with antigens from HSV-1-transformed L cells and transformed-cell antiserum. These two antigens were shown to be present in all clonal lines of HSV-1-transformed cells examined, six L cell lines and one HeLa cell line. Therefore, we conclude that transformation of cells by HSV-1, which is known to be associated with acquisition of viral thymidine kinase, must also be associated with the presence of these two antigens. We performed experiments showing that there are species of HSV-related antibody in HSV-transformed cell antiserum that could not be absorbed out with antigens from HSV-infected L cells. Antibodies present in lytic antiserum were completely removed by antigen preparations from cells lytically infected with HSV-1. Also, lytic antiserum failed to block HSV-related staining of transformed L cells in a direct immunofluorescence test. These results are compatible with one of two notions: either (i) certain genes are expressed during transformation that are not expressed during lytic infection, or (ii) these genes are expressed to a much more reduced extent during lytic infection than in transformed cells.


Anatomy of herpes simplex virus DNA. II. Size, composition, and arrangement of inverted terminal repetitions. Electron microscope studies on self-annealed intact single strands and on partially denatured molecules show that herpes simplex virus 1 DNA consists of two unequal regions, each bounded by inverted redundant sequences. Thus the region L (70 percent of the contour length of the DNA) separates the left terminal region a1b from its inverted repeat b'a'1, each of which comprises 6 percent of the DNA. The region S (9.4 percent of DNA) separates the right terminal region cas (4.3 percent of the DNA) from its inverted repeat a'sc'. The regions of the two termini which are inverted and repeated itnernally differ in topology. Thus, cas is guanine plus cytosine rich, whereas only the terminal 1 percent of the a1b region, designated as subregion a1, is guanine plus cytosine rich.


Effect of DNA-negative and maturation-defective conditions on accumulation of functional messengers for T4 bacteriophage-specific dihydrofolate reductase and deoxynucleoside monophosphate kinase. Messengers for T4 phage-specific deoxynucleoside monophosphate kinase overaccumulated in nonpermissive hosts infected with amber-defective viruses that displayed either the DNA-negative or maturation-defective phenotype. Under both conditions, however, transcription of functional messengers for dihydrofolate reductase followed essentially normal kinetics.


Experience with the thoraco-abdominal incision for nephroblastoma in children less than 3 years old. Our results with the routine use of a modified thoraco-abdominal incision in the treatment of patients with nephroblastoma are reported herein. We have used this incision successfully in 5 patients less than 3 years old and are impressed with the improved exposure. In our limited experience the thoracic extension has not adversely altered the operative or postoperative course.


A review of 100 renal and perinephric sonograms with anatomic diagnoses. Sonography is a non-invasive safe, convenient, quick and pain-free diagnostic modality, which can differentiate fluid masses from solid tumors and give a 3-dimensional representation of organs and masses in the retroperitoneum by biplanar sections. Sonography depends on the fact that high frequency sound has many characteristics of light and is, in fact, a sophisticated method of transillumination. A review of the first 100 consecutive patients who had sonography and anatomic diagnosis of the renal or perinephric abnormality shows the versatility and potentials of sonography. Of these 100 sonograms of renal and perinephric abnormalities 92 were correct by anatomic diagnosis.


Beware of the normal excretory urogram.

Beware of the normal excretory urogram. Excretory urography is not an infallible diagnostic procedure and should not be relied upon exclusively. A normal study must never result in the rejection of additional data derived from history, physical examination and laboratory tests. Two case histories are presented to underscore this caveat. Excretory urography must be supplemented additional radiographic evaluations when the conventional study fails to completely explain the clinical situation.


A serologic survey of pronghorns in Alberta and Saskatchewan, 1970-1972. To determine the exposure of free-ranging pronghorns (Antilocapra americana Ord) to selected pathogens, serum samples were obtained from 33 live-trapped animals from southwestern Saskatchewan in 1970, and from 26 and 51 animals from southeastern Alberta, in 1971 and 1972, respectively. Antibodies were found to the agents of parainfluenza 3, bovine virus diarrhea, eastern and western encephalomyelitis, infectious bovine rhinotracheitis and the chlamydial group. No serologic reactors were found to the agents of bluetongue, epizootic hemorrhagic disease, brucellosis, or leptospirosis (4 serotypes).


Herpersvirus strigis: host spectrum and distribution in infected owls. Herpesvirus strigis, a new species of the genus Herpesvirus, is a pathogen for several species of owls in the order Srigiformes. Natural infection has been observed in the Eagle Owl (Bubo bubo L.), Long-eared Owl (Asio otus L.) and Snowy Owl (nyctea scandiaca L.) In addition the Little Owl (Athene noctua Scopolic) and Tengmalms Owl (Aegolius funereus L.) was experimentally infected. On the other hand the Tawny Owl (Strix aluco L.) and Barn Owl (Tyto albo Scopoli) proved resistant to a massive experimental infection. Of representatives from nine other orders of birds and mammals, only the Old World Kestrel (Falco tinnunculus L.) was found susceptible to this virus. Distribution of viral antigen in various organs of infected owls, as determined by immunofluorescence and by quantitative virus assay, was in accordance with the occurrence of macroscopic and microscopic lesions.


An outbreak of a hemorrhagic disease in white-tailed deer in Kentucky. In 1971, an outbreak of a hemorrhagic disease occurred in captive and free-ranging white-tailed deer (Odocoileus virginianus) in Mammoth Cave National Park, Kentucky, Clinical signs and gross pathological lesions were consistent with those of epizootic hemorrhagic disease and bluetongue, as were serological and histopathological findings for samples sent to other laboratories. The infection rate among the 104 captive deer was 88-92%, and that among the free-ranging Park deer appeared to be similar. Mortality was negligible in the Park deer, but 65 (62%) of the captive deer died. The deaths were bimodally distributed over a 36-day period, and the mortality rate decreased from 97-100% for deer clinically ill during the first 17 days of the outbreak to 58% for deer first exhibiting clinical signs on day 16 or later. Mortality was equal in males and females, but less in yearlings than among fawns or adults. Winter mortality among survivors of the initial outbreak was associated with low ambient temperatures and sometimes fungal and bacterial abscesses, possibly sequelae or complications of the hemorrhagic disease. The pregnancy and birth rates among surviving does appeared to be normal.


Cytomegalic inclusion disease in the east African headgehog. Cytomegaly with intranuclear inclusion bodies was found in the duct epithelium of the salivary glands of five of ten hedgehogs (Atelerix albiventris). Cytomegalovirus infection was presumed to be the cause.


Fatal pox infection in a rough-legged hawk. Natural pox infection occurred in a free-living rough-legged hawk (Buteo lagopus) in northeastern North Dakota. Gross, histological and electron microscopic findings were typical of pox infection, and characteristic lesions developed in red-tailed hawks (Buteo jamaicensis) but not in great horned owls (Bubo virginianus) following inoculation with case material. Death of the rough-legged hawk was attributed to starvation rsulting from inability to capture prey and to blood loss from foot lesions.


The pathogenicity of bovine strains of foot and mouth disease virus for impala and wildebeest. Impala (Aepyceros melampus) and wildebeest (Connochaetes taurinus) were infected with bovine strains of foot and mouth disease virus by intradermolingual inoculation. No clinical signs developed in the impala but mild atypical lesions developed in the tongues of the wildebeest with generalized spread to one foot in two of the eight animals exposed. All the impala but only some of the wildebeest developed viraemia. No virus could be isolated from any tissues in either species after the 7th day following virus inoculation. Immune response occurred in both species. A field survey revealed few animals of either species with significant antibody titers and no virus 'carriers' were found.


Herpes simplex virus in idiopathic facial paralysis (Bell palsy). Sera from all 41 adult patients with idiopathic facil paralysis (Bell palsy) and 35 (85%) of 41 matched controls who had never had Bell palsy contained antibodies to herpes simplex virus (P smaller than.05). The frequency of antibodies to herpes zoster virus did not differ in patients and controls. A rise in antibody titer, indicating primary herpes simplex virus infection, was not found in these patients. That Bell palsy may be caused by reactivation of herpes simplex virus is suggested by (1) clinical, neurologic, laboratory, and immunologic similarities between idiopathic facial paralysis and known manifestations of reactivated herpes simplex virus infection, and (2) the known neurotropism of herpes simplex virus, including its presence in latent form in the trigeminal ganglia, and parallels with known facial paralysis due to varicella zoster virus, a closely related agent. The presence of antibodies to herpes simplex virus is the only common factor among the patients tested in this study.


Methylxanthine induced small intestinal secretion. Methylxanthines, being potent phosphodiesterase inhibitors, produce increased intestinal cyclic AMP levels and would be predicted to produce increased net intestinal fluid secretion. Their effect when presented to the intestinal lumen, which would be analogous to human ingestion, had not been previously determined. Isolated loops of rat jejunum were perfused with solutions of caffeine and theophylline in vivo. There was a decrease in net fluid absorption in both neonatal and mature animals exposed to theophylline. Mature animals exposed to caffeine developed a prompt secretory response, comparable to thax induced by cholera toxin. The data indicate that methylxanthines are potent intestinal secretagogues when administered intraluminally and suggest that secretory stimulation could be important in the gastrointestinal symptomatology elicited in man by these compounds.


[Glomus tumors of the eyelids (author's transl)]. Two new observations of glomus tumors in the eyelid are reported, one of the angiomatous type, the other of the more adenoid type. The typical symptoms of other than ophthalmological localisations are described. The clinical diagnosis is possible. In the few cases reported by ophthalmologists the diagnosis is made by the histologist.


Multiple Herpesvirus simiae isolation from a rhesus monkey which died of cerebral infarction. A rhesus monkey housed in the New England Regional Primate Research Center for more than 4 yr died after an illness of 19 da. Clinical signs included central nervous system involvement and ulcers on the labial mucosa. Microscopically, the cause of death was established as multiple cerebral infarction. Lesions characteristic of herpesvirus infections were not present in the brain nor in any other tissue; however, Herpesvirus simiae was isolated from oral and anal swabs as well as from tongue and lung tissue. Inoculation of this agent in H simiae antibody negative rhesus monkeys resulted in infection, as indicated by virus isolation, serologic conversion, conjunctivitis, and development of ulcers at the mucoepithelial border, but fatal infection did not develop.


Husbandry of the "nude" mouse in conventional and germfree environments. The "nude" mouse is a unique tool for immunologic studies. Its relatively short life span dictates the application of rigid environmental controls to increase longevity if the mouse is to assume the role of a practical experimental animal. In this paper we discussed the husbandry procedures employed to raise "nude" mice in our facilities under conventional, defined flora, and germfree conditions. Conventional and defined flora mice were raised on laminar flow stay-clean rocks, and germfree "nudes" were housed in self-contained germfree isolators. The major cause of morbidity and mortality among conventional and defined flora "nude" mice was fulminating hepatitis. We presented evidence that the etiologic agent of the disease was mouse hepatitis virus (MHV). Germfree "nude" mice were completely free from viral and bacterial diseases.


Human vascular smooth muscle in culture. Growth and ultrastructure. Mixed primary cultures of endothelial and smooth muscle cells were obtained from human umbilical cord vessels after prolonged collagenase digestion of their luminal surfaces. Morphologically homogeneous populations of vascular smooth muscle were then selectively isolated and subcultured for up to 16 weeks. Ultrastructurally, cultured cells were characterized by the presence of bundles of myofilaments with dense bodies similar to native umbilical vessel smooth muscle. Mature cultures developed a distinctive topographical organization consisting of bands of parallel cells and intertwined, multilayered areas. Elaborate intercellular attachments formed along contiguous cell surfaces. Large amounts of extracellular material accumulated, including amorphous substance, elastic fiber microfibrils, and 250- to 300-A,faintly-banded fibrils. In older cultures, focal proliferation, extracellular material secretion and cellular degeneration produced nodular protrusions somewhat resembling atherosclerotic lesions in vivo. Endothelium and smooth muscle cultured from this readily available human source may provide useful comparative material for pathophysiologic studies of vascular disease.


Prevention of pulmonary vascular changes of chronic alveolar hypoxia by inhibition of angiotensin I-converting enzyme in the rat. To test the hypothesis that the renin-angiotensin system is involved in the development of the pulmonary vascular patholigic changes of chronic alveolar hypoxia, two groups of rats were exposed to 0.5 atm. for 21 days. One group received SQ 20,881 (2 mg. per kg.) every 8 hours subcutaneously and the second group received normal saline. A third group of rats was maintained at normobaria. Rats receiving SQ 20,881 had significantly less pulmonary arterial hypertrophy and right ventricular hypertrophy than hypobaric animals. SQ 20,881-treated rats showed a significant increase in adrenal weight but a marked reduction in the thickness of the zona glomerulosa, as compared with the other groups. Our findings indicate that angiotensin II is necessary in the development of the pulmonary vascular structural changes of chronic alveolar hypoxia.