Natural antibody in mammary tumor virus-infected mice that reacts with intracytoplasmic A particles of mouse mammary tumors. By an indirect immunofluorescence technique with prolonged serum incubation on murine mammary tumor (MT) slices, 179 of 424 mice examined were found to possess natural serum antibody (antibodies) that reacted with intracytoplasmic A particles (iAp) of MT cells. The immunologic specificity of this antibody was supported by absorption and blocking experiments. Furthermore, a strong similarity was seen between the mouse antibody reaction on various MT and the fluorescence pattern of rabbit anti-iAp antiserum on these tumors. In female mice, incidence and geometric mean titers of the antibody in part were correlated to the spontaneous MT frequency of the mouse strains examined. Some mice of the strains XVII/Bin and CBA/BinfXVII/Bin, hitherto regarded as "free" of the mouse mammary tumor virus (MuMTV), also contained anti-iAp antibody in their sera. In contrast to MuMTV)-producing CBA/Bin micethese animals did not possess detectable spontaneous antibody reacting with MuMTV-B particles. Therefore, hypothetically, the antibody response in these mice might be induced by incomplete MuMTV expression. In the strain CBA/Bin, females 4 months old and older possessed the antibody in significantly higher geometric mean titers when compared to 4-week-old female mice. The history of lactation seemed to have no influence on the titer of antibody. In the comparatively high MT strains CBA/Bin and C3H/Bin, adult (4-month-old) females had the antibody in significantly higher levels when compared to age-matched males.


Spontaneous lymphosarcoma arising in a nude mouse: characterization in vivo and in vitro. A lymphosarcoma spontaneously arising in a nude mouse and a continuous cell line (NML-1) derived from it are described and compared. The primary tumor and a transplantable tumor line from it were composed of lymphoid cells, with no C-type viral particles seen by electron microscopy. The culture line was composed of cells with morphologic and functional properties of macrophages; budding C-type particles were abundant. The cells in the tumors produced in nude mice by injection of the NML-1 cells also resembled macrophages morphologically rather than lymphocytes; however, by electron microscopy, no C-type particles were seen. The findings suggest some type of in vivo suppression of complete expression of the virus.


Effects of lead and other inhibitors on the activation of K+-dependent p-nitrophenylphosphatase activity by glycine. The in vitro and in vivo effects of lead on the activation of K+-dependent p-nitrophenylphosphatase activity by glycine were investigated in plasma membranes of male Wistar rat livers. This activation was markedly inhibited by lead in vitro (0.1-5.0 mM) and in vivo (intraperitoneal injection of lead at 4, 8, or 20 mg per 100 g body weight). On the other hand, the in vitro activation of this K+-dependent activity by glycine was about 77% inhibited by ouabain (10 micronM), but about 100% inhibited by oligomycin (20 micronM).


Cell-free translation of RNA synthesized in vitro by a transcribing  nucleoprotein complex prepared from purified vesicular stomatitis virus. The RNA species synthesized in vitro by a transcribing nucleoprotein (TNP) complex of vesicular stomatitis virus (VSV) were translated with high efficiency in a fractionated cell-free system derived from reticulocytes. The use of TNP complexes isolated from VSV Indiana, VSV New Jersey, and Chandipura viruses showed that in each case the predominant polypeptides synthesized had electrophoretic mobilities identical to their virion N, NS, and M polypeptides in proportions reflecting those found in infected cells rather than purified virions. A minor polypeptide corresponding to unglycosylated polypeptide G was also observed, but the in vitro synthesis of polypeptide L was not detected. The addition of RNase inhibitor to transcription mixtures markedly increased the rate of RNA synthesis. Furthermore, the messenger activity of the RNA was significantly enhanced. The inclusion of S-adenosyl L-methionine during transcription substantially increased the messenger activity of the product RNA, suggesting a requirement for methylation. Fractionation by oligodeoxythymidylic acid-cellulose chromatography revealed that the RNA required a polyadnylic acid tract for messenger activity.


Distinguishable transformation-defective phenotypes among temperature-sensitive mutants of Rous sarcoma virus. Eight transformation-defective, temperature-sensitive (ts) mutants of the Prague strain of Rous sarcoma virus, subgroup A, have been isolated after mutagenesis with 5-bromodeoxyuridine followed by selection on the basis of focus tests. Five of these mutants, ts GI201, GI202, GI203, GI204, and GI205, exhibit properties like most previously reported isolates in that they show a temperature-sensitive response to each of a variety of transformation-specific parameters tested. Interestingly, GI201, in addition to the temperature-sensitive defect, carries a lesion that was observed as a nonconditional loss of expression of plasminogen activator protease. Three mutants, ts GI251, GI252, and GI253 have been disignated partial transformation-defective (PTD) mutants since they behave as ts mutants according to some tests for transformation and as wild type according to others. These three mutants fail to form foci at the nonpermissive temperature (41 degrees C) and art nontumorigenic in 3-week-old chickens (body temperature, 42 degrees C). The agglutinability by concanavalin A of cells infected with these mutants shows a definite  temperature sensitivity, as do the rate of 2-deoxyglucose uptake and the disappearance of the 250, 000-dalton normal cell glycoprotein (large, external, transformation sensitive [LETS]). Although the PTD mutant-infected cells, unlike cells infected with other transformation mutants, exhibit a cell-bound plasminogen activator protease at the nonpermissive temperature, this activator is not detectable as a free protease in the medium, as it is with wild-type, virus-infected cells. The PTD mutants behave like the wild-type parent in their ability to induce transformed growth properties in the infected cells, i.e., growth beyond normal cell saturation density with or without serum-supplemented medium and growth leading to colony formation in soft-agar- or methyl cellulose-containing suspension media.


Proteins of bovine leukemia virus. I. Characterization and reactions with natural antibodies. The bovine leukemia virus (BLV) was purified from a chronically infected fetal lamb kidney cell line. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of this virus revealed the presence of eight distinguishable viral components with molecular weights ranging from 80,000 to 11,000. The major component is a non-glycosylated protein having a molecular weight of 24,000 (p24). At least three heavier polypeptides were found, one of them representing a glycoprotein (gp 60). In addition, four minor polypeptides with respective molecular weights of 19,000, 16,000, 13,000, and 11,000 were identified. In a complement fixation assay using naturally occurring antibodies of a leukemic cow, four polypeptides, which included gp 60, p35, p24, and p16, were found to be reactive.


RNA synthesis of vesicular stomatitis virus. VII. Complete separation of the mRNA's of vesicular stomatitis virus by duplex formation. Full-length virion RNA and complementary mRNA's of vesicular stomatitis virus can be annealed to each other, digested with RNases, and then separated as five unique duplex RNA molecules on polyacrylamide slab gels. Similar RNA duplexes were detected whether mRNA or virion RNA was the radioactive component and whether the mRNA was synthesized in vitro or in vivo. The sharp banding pattern of these RNA molecules was dependent on treatment with RNase T2, suggesting that removal of poly(A) is necessary. Identification of the coding region contained in each RNA duplex was based on their previous identification as single-stranded mRNA on formamide-containing, polyacrylamide gels. Because the two smallest mRNA'S had not been previously separated, their identification was based on their in vitro transcriptional gene order. In the order of increasing mobilities on the slab gels, the RNA duplexes are identified as the hybrid of the region of the genome RNA hybridized to the complementary mRNA coding for the large protein, the glycoprotein, the nucleocapsid protein, the core-associated NS protein, and the matrix protein (L,G,N,NS, and M). Several lines of evidence support the presence of undegraded complete mRNA, excluding poly(A), in these RNA duplexes. Also, the two smallest mRNA's, separated by duplex formation, were denatured, and their individual oligonucleotide fingerprints were determined. From chemical length determinations, the molecular weights of the mRNA, minus poly(A), are 2.78 X 10(5) and 2.5 X 10(5), respectively, for the mRNA's of the NS and M proteins.


Characterization of vesicular stomatitis virus mRNA species synthesized in vitro. The smallest size class of mRNA (12S) synthesized in vitro by the virion-associated RNA polymerase of vesicular stomatitis virus contains two mRNA species of similar molecular weight that code for the viral M and NS proteins. The resolution of these mRNA species was achieved by converting them to duplexes by annealing with the genome RNA, followed by RNase T2 treatment and separation in a polyacrylamide gel. Using this separation technique, the mRNA's were identified by comparing the relative resistance of their syntheses to UV irradiation of the virus. The molecular weights of these two mRNA species calculated as duplex RNAs were smaller than expected. The possible reasons for this discrepancy are discussed.


Immunological reactivity of antisera to sodium dodecyl sulfate-derived polypeptides of polyoma virions. A study was undertaken to produce antisera to sodium dodecyl sulfate-derived polyoma virion polypeptides. With the use of this antisera, it was possible to detect, by immunofluorescence, cytoplasmic synthesis of V1, V2, and V3 polypeptides at 18 h postinfection and subsequent transport to the nucleus by 22 h postinfection. Anti-V1, anti-V2, and anti-V3 sera did not react with intact virions in an immunodiffusion assay, nor did they possess hemagglutination inhibition or viral neutralization activity. Antiserum produced against the four host histone polypeptides (V4 through V7) demonstrated immunofluorescence when reacted with polyoma-infected cells but not with uninfected cells. Antihistone serum was also capable of neutralizing viral infectivity, inhibiting hemagglutination and reacting with whole virions in an immunodiffusion assay.


Localization of two cellular forms of the vesicular stomatitis viral glycoprotein. Two cell-associated forms of the glycoprotein (G) of vesicular stomatitis virus, termed G1 and G2, have been resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. G1 has the higher electrophoretic mobility, but both forms migrate more slowly than G protein synthesized in a wheat germ cell-free system (G0), which presumably is the unglycosylated form. G1 is a kinetic precursor of the G2 form, and the apparent cause of the electrophoretic difference between the two species is the presence of N-acetylneuraminic acid on the G2 form. Conversion of G1 to G2 occurs 10 to 20 min prior to the appearance of the G2 form of the protein on the cell surface. This suggests that the G protein may be completely glycosylated several minutes prior to its migration to the cell surface and that glycosylation is not the limiting step in its maturation. No glycoprotein comigrating with G0 can be detected in the infected cells, even after 5-min labeling periods; this suggests that partial clycosylation of G occurs concomitantly with or immediately after its synthesis.


Separate pathways of maturation of the major structural proteins of vesicular stomatitis virus. Cell fractionation and protein electrophoresis were used to study the intracellular sites of synthesis and intermediate structures in the assembly of the virion proteins of vesicular stomatitis virus. Each of the three major virion proteins assembled into virions through a separable pathway. The nucleocapsid (N) protein was first a soluble protein and later incorporated into free, cytoplasmic nucleocapsids. A small amount of N protein was bound to membranes at later times, presumably representing either nucleocapsids in the process of budding or completed virions attached to the cell surface. The matrix (M) protein also appeared to be synthesized as a soluble protein, but was then directly incorporated into membranous structures with the same density as whole virus. Very little M protein was ever found in membranes banding at the density of plasma membranes. The M protein entered extracellular virus very quickly, as though it moved directly from a soluble state into budding virus. In contrast, the glycoprotein (G) was always membrane bound; it appeared to be directly inserted into membranes during its synthesis. Glycosylation of the G protein was completed only in smooth membrane fractions, possibly in the Golgi apparatus. After a minimum time of 15 min following its synthesis, G protein was incorporated into the surface plasma membrane, from which it was slowly shed into virions. These multiple processing steps probably account for its delayed appearance in virus. From this work it appears that the three major structural proteins come into the surface budding structure through independent pathways and together they coalesce at the plasma membrane to form the mature virion.


Analysis of the defects of temperature-sensitive mutants of vesicular stomatitis virus: intracellular degradation of specific viral proteins. The metabolism of viral RNA and proteins has been studied in cells infected with temperature-sensitive mutant strains of vesicular stomatitis virus. Certain viral proteins encoded by the mutant strains, usually the putative mutant protein for the assigned complementation group, were shown to be degraded more rapidly at the nonpermissive temperature than were the wild-type proteins. Group III mutants (tsG33, tsM301) encode M proteins which are degraded three- to fourfold faster than the wild-type protein. This defect cannot be fully rescued by coinfection with wild-type virus, and thus the defect appears to be in the M protein itself. Mutants tsM601 (VI) and tsG41(IV) encode N proteins which are degraded much faster than the wild-type protein and also share the property of being defective in replication of viral RNA, suggesting a correlation between these phenotypic properties. Furthermore, the L proteins of tsG11(I) and tsG13(I) are more labile than the wild-type protein at the nonpermissive temperature. The G protein of tsM501(V) did not undergo the change in electrophoretic mobility previously shown to be the result of sialylation, suggesting that it is defective in maturation or glycosylation at the nonpermissive temperature. Three of the mutants previously isolated in this laboratory, tsM502(V), tsM601(VI), and tsM602(VI), were shown to be defective in viral RNA synthesis at the nonpermissive temperature. Mutant tsM601(VI) was defective mainly in viral RNA replication, whereas tsM502(V) appeared to be totally defective for viral RNA transcription and replication at the nonpermissive temperature.


Maturation of viral proteins in cells infected with temperature-sensitive mutants of vesicular stomatitis virus. Maturation of viral proteins in cells infected with mutants of vesicular stomatitis virus was studied by surface iodination and cell fractionation. The movement of G, M, and N proteins to the virion bud appeared to be interdependent. Mutations thought to be in G protein prevented its migration to the cell surface, allowed neither M nor N protein to become membrane bound, and blocked formation of viral particles. Mutant G protein appeared not to leave the endoplasmic reticulum at the nonpermissive temperature, but this defect was partially reversible. In cells infected with mutants that caused N protein to be degraded rapidly or prevented its assembly into nucleocapsids, M protein did not bind to membranes and G protein matured to the cell surface, but never entered structures with the density of virions. Mutations causing M protein to be degraded prevented virion formation, and G protein behaved as in cells infected by mutants in N protein. These results are consistent with a model of virion formation involving coalescence of soluble nucleocapsid and soluble M protein with G protein already in the plasma membrane.


Chromatin-like structures obtained after alkaline disruption of bovine and human papillomaviruses. Four low-molecular-weight polypeptides migrating like H2a, H2b, H3, and H4 calf liver histones were detected by sodium dodecyl sulfate-acrylamide gel electrophoresis of highly purified preparations of bovine papillomavirus (BPV) and human papillomavirus (HPV). Complexes of these polypeptides and viral DNA were isolated by agarose-gel filtration of the alkaline disruption products of both viruses. When observed under the electron microscope, these complexes appeared as circular structures composed of nucleosomes with a diameter of about 8.0 nm interconnected by a naked DNA filament. The maximal frequency of nucleosomes per molecule was 30 for both viruses, corresponding to a condensation ratio of the viral DNA of 2.5.


Human papillomavirus DNA: physical mapping of the cleavage sites of Bacillus amyloliquefaciens (BamI) and Haemophilus parainfluenzae (HpaII) endonucleases and evidence for partial heterogeneity. The DNA of human papillomavirus (HPV) obtained from a pool of plantar warts is cleaved by bacillus amyloliquefaciens (BamI) and Haemophilus parainfluenzae (HpaII) restriction endonucleases at one and four specific sites, respectively. These sites were localized on the previously established cleavage map of HPV DNA, using the Hind, HindIII, HpaI, and EcoRI endonuclease restriction sites as reference. The four HpaII sites were mapped, clockwise, at 1.4, 41.1, 44.3, and 52.8% of the genome length from the unique BamI cleavage site taken as point zero. The HpaII site mapped at 1.4% of the genome length was absent in 40 to 50% of the molecules, thus showing a genetic heterogeneity of HPV DNA.


Contact-inhibited revertant cell lines isolated from simian virus 40-transformed cells. VII. Serum detachment-resistant revertant cells. Flat revertant cells of simian virus 40-transformed mouse fibroblasts have been isolated on the basis of resistance to a selective detachment procedure. The revertants are generally similar to those isolated by other procedures.


Induction of deoxypyrimidine kinase activity in human embryonic lung cells infected with varicella-zoster virus. Deoxypyrimidine kinase (deoxythymidine [TdR] kinase and deoxycytidine kinase) activity was induced in human embryonic lung cells after infection with varicella-zoster virus (VZ virus). Increased enzyme activity was also produced by using cell-associated virus as inoculum instead of cell-free virus. Anti-VZ virus serum inhibited both the appearance of cytopathic effect and the induction of enzyme activity. The induced TdR kinase activity was more thermostable than that induced by herpes simplex virus type 1. Also, the TdR kinase activity of VZ virus-infected cells was inhibited by dTTP less than in mock-infected cells and more than in herpes simplex virus type 1-infected cells.


Pseudotypes of vesicular stomatitis virus with the mixed coat of reticuloendotheliosis virus and vesicular stomatitis virus. Vesicular stomatitis virus (VSV) forms pseudotypes with envelope components of reticuloendotheliosis virus (REV). The VSV pseudotype possesses the limited host range and antigenic properties of REV. Approximately 70% of the VSV, Indiana serotype, and 45% of VSV, New Jersey serotype, produced from the REV strain T-transformed chicken bone marrow cells contain mixed envelope components of both VSV and REV. VSV pseudotypes with mixed envelope antigens can be neutralized with excess amounts of either anti-VSV antiserum or anti-REV antiserum.


Factors affecting composition and thermostability of mengovirus virions. The composition of mengovirus virions produced by infected cells varies with the incubation temperature. Virons produced at 37.0 or 39.5 degrees contain four major polypeptides (alpha, beta, gamma, and delta) and one minor polypeptide (beta'). Virons produced at 31.5 degrees C contain two additional polypeptides (D1 and E). The virions of two temperature-sensitive (ts) and thermolabile mutants of mengovirus (ts25 and ts88) contain an increased amount of polypeptide beta', with a corresponding decrease in polypeptide beta when compared with the wild-type mengovirus.


Amount and distribution of virus-specific sequences in giant RNA molecules isolated from polyoma-infected mouse kidney cells. A two-step hybridization with polyoma DNA was used to study the composition of giant RNA molecules synthesized in mouse kidney cells late in productive infection by polyoma virus. Giant molecules longer than a complete transcript of the polyoma genome were purified from cells that had been pulse-labeled for 30 min with [3H]uridine and annealed, under mild conditions (50% formamide, 37 degrees C), with polyoma DNA loaded on nitrocellulose filters. Hybridized RNA (6 to 7% of the entire population of 3H-labeled molecules and up to 15% of the molecules containing polyadenylic acid [poly(A)]] was eluted and annealed a second time with polyoma DNA under more stringent conditions. In this second step, 75% of the 3H-labeled RNA formed an RNase-resistant hybrid. Under the same conditions, complementary RNA hybridized with polyoma DNA to a maximal extent of 80%. Since the difference between 75 and 80% is within the experimental error of the hybridization assay, it is inferred that the giant molecules selected by the first hybridization may consist entirely of virus-specific sequences or contain, at the most, a minor fraction of nonviral sequences. To examine the possibility that such nonviral sequences are clustered at the 3'-terminus of these molecules, poly(A)+ giant RNA, which had not been preselected by hybridization with polyoma DNA, was fragmented by a limited alkaline hydrolysis. Fragments linked to the poly(A) segment were separated from the rest of the cleavage products. A one-step hybridization with polyoma DNA revealed that both fractions contain 8 to 10% of virus-specific sequences. These results indicate that the 3'-termini of the poly(A)+ polyoma-specific giant RNA molecules consist of viral rather than nonviral sequences.


Regulation of Sendai virus transcription: evidence for a single promoter in vivo. The synthesis of Sendai virus RNA species was examined after UV irradiation of cells late in infection. Compared with the inactivation of 50S genomic RNA synthesis, the synthesis of the group of mRNA species that sediments at 18S was inactivated at an average rate consistent with a process of sequential transcription from a single promoter. The rates of inactivation of the synthesis of individual mRNA's separated by polyacrylamide gel electrophoresis confirmed this and, with the aid of additional data, suggested that the order of genes in the Sendai virus genome is: 3'-NP-Fo-M-P-HM-L-5'.


Cell killing by simian virus 40: impairment of membrane formation and function. Simian virus 40 infection of the CV-1 line of green monkey kidney cells results in the release of mitochondrial malic dehydrogenase as early as 24 h. Released malic dehydrogenase is detected in the cytoplasm prior to its appearance in the overlay medium. Infected cells lose the ability to consume oxygen between 48 and 56 h, and damage to the elctron transport system is indicated. Nevertheless, cellular ATP levels remain high as late as 72 h. Infection leads to a stimulation of membrane phospholipid synthesis, which reaches a peak at about 32 h. This is followed by a severe decline in new membrane synthesis, which correlates in time with the release of cytoplasmic lactic dehydrogenase into the overlay media. Lactic dehydrogenase release precedes the accumulation of trypan blue-stainable cells by about 6 h. Infection had no effect on the turnover of prelabeled membrane phospholipids. An early simian virus 40 mutant, tsA58, and a late mutant, tsB11, are both less effective than wild-type virus at causing reduced levels of phospholipid synthesis, enzyme release, and the accumulation of trypan blue-stainable cells. Another late mutant, tsB8, is similar to wild-type virus in these respects. At 64 h, there is no detectable cell-associated lactic dehydrogenase and nearly all the cells are trypan blue stainable. Nevertheless, at concentrations of deoxyglucose in the medium below the transport Km, deoxyglucose uptake was similar in infected and control cultures. With higher concentrations of deoxyglucose in the medium, uptake by the infected cultures exceeded that by the control cultures.


Quantitation of herpes simplex virus type 1 RNA in infected HeLa cells. We have quantitatively analyzed the size and amount of herpes simplex virus (HSV)-specific RNA synthesized in HeLa cells using DNA and RNA excess hybridization. At 2 h after infection (early), transcripts from 20% of the total HSV DNA are present on polyribosomes as poly(A+) RNA. At this time, viral poly(A+) RNA comprises 60 to 75% of the newly synthesized poly(a+) mRNA on polyribosomes. By 6 h after infection (late), poly(A+) HSV RNA transcribed from 35 to 40% of the viral DNA is found on polyribosomes. These viral poly(A+) transcripts comprised as much as 90% of newly synthesized poly(A+) mRNA and are measurably larger than viral poly(A+) transcripts isolated early. Some but not all of this size difference is due to the fact that the poly(A) tails on early transcripts are shorter than those found on transcripts made late. Even late after infection, a small but readily measurable amount of cellular poly(A+) RNA is still being made and entering polyribosome complexes. In the nucleus, late after infection, poly(A+) HSV RNA is complementary to 50% of the total HSV DNA. Both early and late after infection, total nuclear viral transcripts are, on the average, larger than viral transcripts found on polyribosomes; however, nuclear HSV poly(A+) RNA is not measureably larger than the corresponding cytoplasmic viral poly(A+) sequences at either time. A major portion (30 to 40%) of the polyribosomal HSV RNA made either early or late after infection is not polyadenylated. This HSV poly (A-) RNA is transcribed from the same sequences as HSV poly(A+) RNA but, when labeled and isolated either early or late after infection, both nuclear and polyribosomal viral poly(A-) RNA molecules sediment faster in sucrose-formaldehyde gradients than their polyadenylated counterparts.


Divergence of baboon endogenous type C virogenes in primates: genomic viral RNA in molecular hybridization experiments. RNA purified from two related RNA tumor viruses, one isolated from a baboon, Papio anubis, and the second from cultured blood leukocytes of a patient with acute myelogenous leukemia, was labeled with 125I and hybridized to DNA from different primates. RNA from both viruses showed maximum sequence homology with genes in baboons and little homology with genes of humans. The results confirm earlier suggestions that both viruses originated by transcription of baboon virogenes, and that one was transmitted to humans in nature. Hybridization of the viral RNA to cell DNA followed complicated kinetic patterns, indicating the presence of both repeated and infrequent virogene elements. This conclusion was verified in experiments using varied DNA:RNA ratios. It is proposed that virogenes, though composed of genes repeated 10 times or more, consist of some sequences more preferentially conserved than others. The non-uniformity of virogene sequence conservation limits the use of viral probes in studies concerning certain aspects of virogene evolution.


Homogeneity and complexity of avian oncornavirus proviral DNA determined by molecular hybridization. The homogeneity of DNA complementary to the 35S RNA subunit of avian myeloblastosis virus (AMV) has been demonstrated by single or multistep hybridization. For multistep hybridizations, 35S AMV RNA was preselected for its ability to hybridize either to unfractionated leukemic DNA or to leukemic DNA enriched for unique or for reiterated sequences. These experiments indicate that the viral genome is complementary to DNA sequences with a low reiteration frequency. Competition experiments confirm the absence of fast-hybridizing sequences in viral DNA. Computer analyses of the data reveal that there are two to four copies of viral DNA in infected cells.


Mechanism of restriction of ecotropic and xenotropic murine leukemia viruses and formation of pseudotypes between the two viruses. Ecotropic and xenotropic murine leukemia viruses (MuLV's) constitute separate interference groups; within each group there is cross-interference, but between the groups there is no detectable interference. Interference is manifest against pseudotypes in which the vesicular stomatitis virus genome is contained within the coat of one of the murine leukemia viruses. The pseudotypes display the cell specificity of the leukemia viruses: pseudotypes with an ecotropic MuLV coat infect mouse cells but not rabbit or mink cells; pseudotypes with a xenotropic MuLV coat infect rabbit or mink cells well but mouse cells very poorly. Efficient pseudotype formation also occurs between the two MuLV classes, and both the interference patterns and the cell specificity of these pseudotypes are entirely determined by their envelope. Using these pseudotypes, ecotropic MuLV infection could be established in xenogeneic cells, and the resulting progeny could be scored by using a conventional XC cell assay. Also, xenotropic MuLV infection could be established in a mouse cell, showing that no absolute intracellular barrier against xenotropic virus growth exists in murine cells. The major barriers against both xenotropic and ecotropic MuLV therefore are cell surface barriers. Xenogeneic cells probably lack receptors for ecotropic MuLV, but murine cells may either lack receptors for xenotropic MuLV or have receptors that are blocked by endogenous expression of the glycoprotein of endogenous xenotropic MuLV.


Natural immunity in mice to the envelope glycoprotein of endogenous ecotropic type C viruses: neutralization of virus infectivity. The ability of naturally immune mouse sera to neutralize ecotropic AKR murine leukemia virus (MuLV) was examined by using unfrozen virus preparations harvested for 1 h. In this assay several mouse sera significantly and consistently neutralized MuLV infectivity. The ability of these sera to neutralize was correlated with the presence of antibodies against MuLV detectable in a radioimmune precipitation assay using radioactively labeled intact virions. This neutralization was specific, in that either N- or B-tropic viruses, but not Friend MuLV, were neutralized. In addition, neutralization could be abrogated with purified AKR MuLV gp71 at concentrations that do not interfere with virus infectivity but could not be abrogated with Rauscher MuLV gp71. Neutralizing activity could be removed by absorption with intact AKR MuLV, but not by absorption with Friend MuLV, a BALB/c xenotropic virus, or with NZB xenotropic virus. All the neutralizing activity of (B6C3)F1 mouse sera was associated with the immunoglobulin G fraction.


Isolation of the mouse mammary tumor virus sequences not transmitted as germinal provirus in the C3H and RIII mouse strains. Radioactive 60-70S RNA from the mouse mammary tumor virus (MMTV) produced by the C3H mouse mammary tumor cell line (Mm5mt) hybridized to a greater extent, and at a lower Cot1/2 value, to the DNA of C3H mammary tumor cells than to the DNA of C3H liver cells. The 125I-labeled MMTV (C3H) 60-40S RNA was annealed to a vast excess of DNA from C3H livers, and single-stranded RNA was eluted from hydroxylapatite and recovered. This "recycled RNA" did not hybridize to the DNA of the apparently normal organs tested from normal or from mammary tumor-bearing C3H mice, but hybridized extensively to both the DNA from the C3H mammary tumor cell line and the DNA from spontaneous C3H mammary tumors. This hybridization could be competed out by the addition of unlabeled MMTV 60-70S RNA but was unaffected by the addition of unlabeled 60-70S RNA of C3H type C virus. Similar experiments were conducted with the RIII mouse strain. We therefore report on the isolation of the sequences of the RNA genomes of the MMTVs from C3H and RIII mice that are transmitted by some mechanism other than via the germ line. These studies further define the differences, via molecular hybridization, between the MMTV-S and the MMTV-L in both C3H and RIII mice.


Evidence that herpes simplex virus DNA is transcribed by cellular RNA polymerase B. In herpes simplex virus type 1 (HSV-1)-infected HEp-2 cells, amanitin added before or at various times after infection always reduced viral multiplication. Also, the three waves of transcription of HSV-1 DNA, which led to the synthesis of alpha, beta-, and gamma-polypeptides, were all sensitive to amanitin in HEp-2 cells, and the amanitin-sensitive RNA polymerase activities of isolated nuclei were equally sensitive to the inhibitor before and during the infection. On the contrary, HSV-1 DNA transcription was totally unaffected by amanitin in AR1/9-5B cells, a mutant subline of CHO cells that possesses an amanitin-resistant RNA polymerase B. Together, these results strongly suggest that HSV-1 DNA utilizes for its transcription a polymerase undistinguishable from host cell RNA polymerase B with respect to its sensitivity to amanitin.


Klippel-Trenaunay syndrome associated with urinary tract hemangiomas. Genitourinary hemangiomas in 2 children with Klippel-Trenaunay syndrome are reported. The presence of cutaneous hemangiomas should alert urologists to the possibility of urinary tract hemangiomas inpatients with gross hematuria. Transurethral manipulation of bladder hemangiomas should be avoided because of the danger of excessive hemorrhage, the treatment of choice being partial cystectomy.


Nephroblastoma with right atrial extension: preoperative diagnosis and management. A 14-year-old black boy, with a large nephroblastoma of the right kidney, had preoperative identification of inferior vena cava and right atrial involvement. Renal arteriography revealed linear arterial channels in the anatomic distribution of the inferior vena cava and venography revealed total caval occlusion and a right atrial mass. Radical excision, using a cardiopulmonary bypass, resulted in a 6-month postoperative survival.


Studies on serum lipoproteins in renal diseases. Serum lipoproteins of 25 patients with uncomplicated renal diseases were studied with polyacrylamide gel block electrophoresis. (1) In the nephrotic syndrome (or nephrotic phase of nephritis), marked increases of chylomicrons, pre-beta lipoprotein or beta lipoprotein and decreases of alpha lipoproteins were detected in the most of the patients. Qualitative change was also frequent as in the glomerulonephritis. (2) In the renovascular hypertension, metabolism of serum lipoproteins was involved also. The principal abnormality was in alpha lipoproteins including the lipid-loaded albumin. Marked increases of alpha lipoproteins, to the level equal to or more than beta lipoprotein, was detected in 2 of 3 patients studied in this period. Surgical correction of abnormal physiology had resulted in a return to a normal lipoprotein profile. (3) In the glomerulonephritis confirmed by biopsies, serum lipoprotein abnormalities were detected more frequently than in the reported past studies as analyzed with the method employed in this study. Qualitative as well as quantitative abnormalities were in beta lipoprotein and alpha lipoproteins in the early and middle phase of the disease process. Gross qualitative change occured frequently. Furthermore, lipoprotein abnormalities in renal diseases were reversible; i.e., when the disease had ameliorated or was corrected surgically, the lipoprotein profile returned to the normal or near-normal profile. In conclusion, the results of the present study indicated that serum lipoprotein disorders are involved in the disease process of three major clinical entities of the renal diseases.


Inhibitory effects of prostaglandin E1 and E2 on cholinergic transmission in isolated canine tracheal muscle. The contractile response of the isolated canine tracheal muscle to the transmural nerve stimulation was depressed by atropine and augmented by physostigmine, indicating that the response was predominantly mediated via the parasympathetic nerve. The contractile response to the transmural nerve stimulation was inhibited by prostaglandin E1 (PGE1) and E2 (PGE2) (10(-7) to 10(-5) g/ml) and the inhibitory action of PGE1 was more potent than that of PGE2. On the other hand, the contractile response of the tracheal muscle to exogenously administered ACh was unaffected by 10(-6) g/ml of PGE1 and PGE2. These findings lend support to the hypothesis that the PGE series, in a manner similar to adrenergic transmission, are involved in a negative feed-back control mechanism for the transmitter release in cholinergic transmission.


Influences of aminophylline and reduction in external Na on the antispasmodic action of isoproterenol in the isolated rat rectum. Effects of aminophylline and the reduction in external Na on the antispasmodic action of isoproterenol were investigated in relation to the mobilization of Ca in the isolated rat rectum. The antispasmodic action of isoproterenol on the phasic contractions by acetylcholine and K both in Ca-free and in Ca-free and Na-poor media was potentiated by treatment with aminophylline, however the antispasmodic action was attenuated by reducing Na in the Ca-free medium. Dibutyryl cyclic AMP inhibited media and the inhibitory action was also potentiated by treatment with aminophylline, while the inhibitory action of dibutyryl cyclic AMP was attenuated by reducing Na in the Ca-free medium. From these findings, it appears that isoproterenol inhibits the release of Ca from storage sites induced by acetylcholine and K via the increase of intracellular cyclic AMP content and that the external Na may play an important role in the Ca release-inhibiting effect of cyclic AMP.


[Effect of heparin on blood lipids under conditions of endo- and exogenous hyperlipemia]. In 15 patients suffering from ischemic heart disease and in 5 healthy individuals (controls) the dynamics of the blood lipids content (NEFA, triglycerides and atherogenic lipoproteins) was followed up on an empty stomach, during a 18-hour fasting, after carbohydrate and fat loadings, before and after intravenous administration of heparin. Data on a quick and abrupt rise of the NEFA level and a falling triglycerides concentration, and also on a certain decline in the content of atherogenic lipoproteins following administration of heparin were obtained. In atherosclerotic patients the development of hypertriglyceridemia in response to introduction of glucose and a prolonged rise in the level of triglycerides following a fat loading were recorded. Intravenous administration of heparin with carbohydrates and fat loads contributes in atherosclerotic patients to a rapid fall of the blood plasma tryglycerides concentration.


[Characteristics of energy metabolism in the myocardium under artificial hypothermia]. The effect of cooling and subsequent rewarming on the tissue respiration of canine hearts was studied during polycomponent ether-oxygen anaesthesia. The tests included the determinations of the activity of the dehydrogenases of the cytrate cycle, the content and activity of chromoproteids, the respiration rate of the mitochondrias on succinate, glutamate and ketoglutarate, the content of glycogen, the activity of the phosphorylases, hexokinase, lactate dehydrogenase, the content of lactate, pyruvate, adenyl nucleotides and creatine phosphate. Significant changes were noted in the content and activity of the above substances, acceleration of mitochondrial respiration, reduced energy regulation of respiration, and decreased amount of the adenyl components. It is suggested that under artificial hypothermia the processes of chromoproteids biosynthesis are enhanced, which results in an increased power of terminal respiration, and conformational rearaangements of the enzymes connected with the membranes occur.


Effect of parathyroid hormone and cyclic adenosine 3',5'-monophosphate on isotonic fluid reabsorption: polarity of proximal tubular cells. Isotomic fluid reabsorption (JV) of rat renal proximal tubules was examined by the shrinking droplet method in combination with simulatneous perfusion of blood capillaries. Sensitivity of JV measurement was improved by using each punctured tubule for control measurements: 1) Parathyroid hormoen (PTH) on the contraluminal cell side reduced JV in a dose-response behavior. The maximal inhibition was achieved at a PTH concentration of 10(-5) M, the half maximal inhibition at a concentration of 3 X 10(-9) M. PTH on the luminal cell side had a small inhibitory effect. 2) Cyclic AMP inhibited JV preferentially when applied to the luminal cell side. On the luminal cell side, both cyclic AMP and dibutyryl cyclic AMP inhibited JV in a similar dose-dependent behavior. Concentrations of both nucleotides as low as 10(-10) M had a definite inhibitory effect. Tested at a high concentration, N6-butyryl cyclic AMP was almost as effective as cyclic AMP. Deoxy cyclic AMP, 5' AMP, cyclic guanosine monophosphate (cyclic GMP), dibutyryl cyclic GMP had no effect. ATP inhibited JV to a very small extent. 3) The reduction of JV after administration of PTH and dibutyryl cyclic AMP was not additive. The similar inhibitory effect of PTH at the contraluminal cell face and of cyclic AMP at the luminal cell face suggests the following sequence of events in the mediation of the action of PTH: 1) activation of adenylate cyclase by PTH in the contraluminal cell membrane, and 2) action of the generated cyclic AMP on the luminal cell membrane. The interaction of cyclic AMP and the luminal cell membrane is initiated at the luminal cell surface.


Hyperparathyoidism: influence of glomerular filtration rate on urinary excretion of cyclic AMP. Urinary cyclic AMP excretion per 24 h or per g creatinine in primary hyperparathyroidism (1 degrees HPT) has been evaluated by several authors with conflicting results. In 50 patients with 1 degrees HPT, 25 patients with secondary (2 degrees) HPT and 35 healthy control persons we determined urinary cyclic AMP per 24 h or per g creatinine. These parameters did not satisfactorily discriminate patients from controls, especially when glomerular filtration rate (GFR) as determined by creatinine clearance was reduced. Since urinary cyclic AMP is derived from plasma by glomerular filtration and from kidney by tubular production-the amount of tubules is reflected by GFR-the cyclic nucleotide was related to GFR. In controls urinary cyclic AMP correlated better with GFR than with creatinine excretion. Additionally, in 45 of 50 patients with 1 degrees HPT and in all with 2 degrees HPT, urinary cyclic AMP/GFR was raised. In 1 degrees HPT serum levels of parathyroid hormone correlated closer with urinary cyclic AMP/GFR than with urinary cyclic AMP/g creatinine. The ratio cyclic AMP/GFR decreased to normal or subnormal values after removal of adenomatous or hyperplastic glands in 1 degrees HPT and during infusion of calcium in 2 degrees HPT. In 50 patients with renal lithiasis caused by diseases other than 1 degrees HPT (anatomical variations, pyelonephritis, immobilization after tetraplegia) the ratio cyclic AMP/GFR was not raised. Urinary cyclic AMP/GFR, therefore, reflects parathyroid hormone excess more reliably than cyclic AMP/g creatinine.


Effect of ethane-1-hydroxy-1,1-diphosphonate (EHDP) on the ultrastructure of parathyroid glands and plasma immunoreactive parathyroid hormone in pregnant cows fed a low calcium diet. The long term (70 days) effects of administering ethane-1-hydroxy-1,1-diphosphonate (EHDP) (4 mg. per kg. per day) on parathyroid function was investigated in pregnant cows fed a low calcium diet. Serum calcium and phosphorus were significantly lower at parturition and postpartum in EHDP-treated cows compared to pregnant control cows fed the low calcium diet. Plasma immunoreactive parathyroid hormone levels were similar prepartum, at parturition, and postpartum in cows administered EHDP and control cows. Immediately available calcium reserves were greater preparation in control cows than in cows receiving EHDP as indicated by a more rapid rate of return of serum calcium toward normal levels following ethylenediaminetetraacetic acid (EDTA)-induced hypocalcemia approximately 10 days prepartum. EHDP-treated cows responded to the hypocalcemic challenge with similar changes in plasma immunoreactive parathyroid hormone levels as in control cows; however, urinary hydroxyproline excretion increased at certain intervals only in control cows. Ultrastructurally, chief cells in parathyroid glands of both groups of cows were in an active stage of the secretory cycle with well developed organelles concerned with hormonela synthesis. Chief cells in cows administered EHDP were degranulated and contained fewer secretory granules in response to the hypocalcemia than those in control cows. Chief cells in EHDP-treated cows often had prominent perinuclear accumulations of microfilaments, scattered vacuolated mitochondria, and lysosomal bodies in the cytoplasm. Thyroid C-cells were densely granulated and thyroid calcitonin content was similar in both groups of cows. The principal defect in calcium homeostasis of EHDP-treated cows appeared to be an impairment both in bone calcium mobilization and bone matrix catabolism in response to the secretion of parathyroid hormone. In vitro uptake of 45Ca by duodenal mucosa and urinary excretion of cyclic adenosine monophosphate were similar in both groups of cows. The ability of the parathyroid glands to synthesize and secrete parathyroid hormone in response to hypocalcemia induced either by EDTA or associated with parturition was not impaired by the administration of EHDP.


Platelet aggregating material in mouse tumor cells. Removal and regeneration. The platelet aggregating principle of two mouse ascites tumors and of their cell-free supernatants released spontaneously has been studied. It was found that the principle disappeared from the cells after trypsin digestion and that part of it was recovered in the cell-free trypsinate. Digested cells regenerated the principle during subsequent incubation by a process requiring protein synthesis. The principle was found to be spontaneously released by intact cells into the medium and sensitive to proteolytic attack. The principle was not present in five varieties of nonneoplastic cells. Since previous work by the authors indicates that the principle is present in numerous other tumor cell lines, its study might reveal it to be an indicator of malignant transformation or malignant progression.


Post-kala-azar dermal leishmaniasis simulating leprosy. (A case report). The case of a 40 years male suffering from post-kala-azar dermal leishmaniasis simulating nodular lepromatous leprosy is reported. In countries where leprosy is endemic, other diseases are not infrequently taken to be leprosy (Schaller, 1971). Dermal leishmaniasis is a common cause of confusion in countries where the condition is endemic (Browne, 1964). Dharmendra and Chatterji (1940) discussed in detail the question of differential diagnosis between Leprosy and Dermal Leishmaniasis. The present communication is concerned with a patient suffering from post-kala-azar dermal leishmaniasis whose skin lesions simulated nodular lepromatous leprosy.


The correlation of plasma membrane microvilli and intracellular cyclic AMP content in a rat epitheloid kidney cell line. Modulation of the intracellular concentration of cyclic AMP has been associated with a regulatory role in cell division, cell morphology, and physical properties of the plasma membrane. Untransformed rat kidney cells in culture exhibit epitheloid morphology, high intracellular cyclic AMP levels, and contact inhibition of growth. Untransformed rat kidney cells transformed with the Kirsten murine sarcoma virus exhibit a low cyclic AMP content, rapid growth rate, and a loss of contact inhibition. Scanning electron microscopy reveals a distinctive difference in the surface structure of the two cell types during Gl of the cell cycle. The surface of the transformed cell is covered with microvilli while its untransformed counterpart is devoid of microvilli. The presence of microvilli can be controlled as a function of temperature by two temperature-sensitive mutants of the Kirsten sarcoma virus (ts6t6 and ts371 cl 5). In the ts6t6 mutant, growth at 32 degrees C results in a low cyclic AMP content and the presence of microville, while growth at 39 degrees C results in a high cyclic AMP content and a decrease in microvilli. The opposite effect is seen with the ts371 cl 5 mutant. Correlation of cyclic AMP content with the presence of microvilli suggests that this surface phenomenon is a function of cyclic AMP concentration.


Quality-of-care assessment. II. Outpatient medical care following hospital dismissal after myocardial infarction. This study was undertaken to compare process and outcome methods of quality assessment of medical care in outpatient office practice. Follow-up care after hospitalization for first acute myocardial infarction was used as the model. One hundred fifty-two patients followed up for a minimum of 2 years comprised the study group. An expert committee of cardiologists and internists in community practice established the process criteria for satisfactory care and predicted outcomes of continuing disability and mortality. Using weighted process criteria and a weighted performance index permitted demonstration of a significant association between process items performed at the first posthospitalization visit and 2-year mortality. A significant association could not be demonstrated between later process of care and outcome at 2 years. The outcome assessment study disclosed that predicted disability and mortality rates compared closely with observed outcomes. However, this method for evaluating the quality of outpatient medical care is weakened because little information is available to provide the basis of prediction of satisfactory outcome rates in complicated cases. Although both the process and outcome methods of quality assessment have short comings, the latter method is recommended because satisfactory outcomes is the essential criterion of quality medical care. Moreover, when process items are not specified outcome assessment maintains the flexibility of individual physician practice. Refinement of satisfactory outcome prediction for common illnesses managed in office practice should be the goal for future studies.


Decreased adenosine cyclic 3',5'-monophosphate phosphodiesterase activity in rat straitum following chronic haloperidol treatment. The possible involvement of cyclic nucleotide phosphodiesterase (PDE) in the supersensitivity to dopamine-receptor agonists after chronic treatment with neuroleptic drugs has been studied. Rats were given haloperidol in the drinking water for 18 days and finally injected i.p. with 10 mg/kg haloperidol. During and after this treatment the low Km form of the cyclic AMP PDE in a 10,000 g supernatant of the striata was reduced. The loss in enzyme activity was associated with a change in the chromatographic behaviour on DEAE-cellulose. The difference between control- and haloperidol-treated rats was most pronounced in the presence of 1 mM ethylene glycol-bis(aminoethylether)tetraacetic acid (EGTA) and was essentially abolished at 1 mM Ca++. This decrease in cyclic AMP PDE may explain some of the supersensitivity to dopamine-receptor agonists observed following chronic neuroleptic treatment.


Combined approach surgery for removal of glomus jugulare tumors. The ideal surgical treatment for glomus jugulare tumors is total removal. Efforts have been made periodically to accomplish this since shortly after this tumor was first identified in the early 1940's. This paper describes a method of removal using a combined approach through the neck and temporal bone, preceded by a course of preoperative irradiation therapy. The early results that have been obtained using this procedure in 10 patients are reported.


Localization of collagenase in human middle ear cholesteatoma. The process of connective tissue breakdown in chronic otitis media is described in the context of recent advances in our understanding of collagen degradation and bone resorption. The significance of the initial step in collagen breakdown, brought about by the action of a specific collagen dissolving enzyme is emphasized in terms of recent studies in other chronic inflammatory diseases characterized by connective tissue breakdown. Bone resorption, a characteristic feature of chronic otitis media, requires the breakdown of collagen, which comprises over 90 percent of bone protein. Evidence in support of collagenase in bone resorption from adjacent tissue (in this case, inflammatory connective tissue) would require identification of the enzyme in cells involved in the inflammatory process adjacent to the resorbing bone. Collagenase was found localized in frozen sections of canal wall skin, middle ear granulation and in cholesteatoma by a specific binding of the enzyme with an antiserum produced against purified human skin collagenase. The antigen antibody complex was labelled with fluorescein. Collagenase appeared in the subepithelial connective tissue of cholesteatoma, granulation tissue from the middle ear and the dermis of canal skin; but was not seen in the keratin layer, epithelium or the epidermal appendages. The enzyme appeared within certain fibroblasts, macrophages and endothelial cells of capillary buds. Collagenase enhanced by chronic inflammation attacks the intact collagen molecule, making it susceptible to further digestion by other proteases that are also products of inflammation. This process brings about resorption of connective tissue and bone.


Activity of phospholipid-synthesizing enzymes in rat liver plasma membranes and the source of biliary lecithin. The potential for the synthesis of phosphatidylcholine by the bile canalicular membrane of the liver cell was assessed by measuring the activity of a number of phospholipid synthesizing enzymes in isolated bile canalicular membrane fractions from rat liver. The activity of these various enzymes was compared to that present in noncanalicular liver cell plasma membranes and in microsomes. The CDP-choline:1,2-diacyl-sn-glycerol-cholinephosphotransferase was virtually absent from the bile canalicular membranes but the specific activities of S-adenosyl-L-methionine:phosphatidyl-ethanolamine N-methyltransferase and acyl-CoA:1-acyl-sn-glycero-3-phosphorylcholine acyltransferase were 11-15% of those found in the microsomes. The bile canalicular membranes also contained detectable acyl-CoA:sn-glycero-3-phosphate acyltransferase activity and the ability to potentiate the Ca++-stimulated exchange of bases between different phospholipids. These findings indicate that the bile canalicular membranes have a very limited capacity for the formation of phosphatidylcholine under the assay conditions employed.


Origin of fatty acids of cholesteryl ester accumulated by Fu5AH cells in culture. The Fu5AH rat hepatoma cell line accumulates cholesteryl ester (CE) upon incubation in medium supplemented with hyperlipemic serum or hyperlipemic serum lipoproteins. This cell line was used to investigate the origin of the fatty acids esterified to cholesterol in intracellular accumulations of CE. The intracellular CE-fatty acid distribution was found to be markedly different from that of the lipoprotein which stimulated the accumulation. Free fatty acids added to the culture medium were found esterified to cholesterol in the cells, demonstrating that cellular esterification contributes to the accumulation of CE. Using a subline of Fu5AH cells containing radioactivity labeled intracellular fatty acids, it was found that about one-third of the fatty acid moiety of CE accumulated by the cells during a 24 hr incubation with hyperlipemic serum was derived from endogenous fatty acids. The drug chloroquine was found to inhibit cellular cholesterol esterification, so that only 4% of CE-fatty acids were derived from endogenous fatty acids. Evidence is presented suggesting a major role for cellular esterification in CE accumulation by Fu5AH cells.


Lipids of cultured hepatoma cells: VIII. Utilization of D-[1-14C] glucose for lipid biosynthesis. Minimal deviation hepatoma 7288C cells (HTC) were incubated in serum-supplemented and serum-free Swim's 77 medium in the presence of D-[1-14C] glucose for 1, 2, 4, 8, 12 and 24 hr. Glucose oxidation to CO2, incorporation into total cell mass, and incorporation into cell and medium lipids were determined. The percentage distribution of total cell lipid radioactivity in individual neutral and polar lipid classes was followed as a function of time. Degradation studies of individual lipid classes were performed to ascertain the percentage of radioactivity in acyl and glycerol moieties. The percentage of D-[1-14C] glucose oxidized to 14CO2, incorporated into cell matter and cell lipids was elevated in cells incubated in serum-free medium as opposed to serum-supplemented medium. The percentage distribution of total cell lipid radioactivity into individual neutral lipid classes from both serum-free and serum-supplemented cultures was as follows: sterols greater than triglycerides greater than free fatty acids greater than sterol esters. The percentage distribution of total cell lipid radioactivity into individual polar lipid classes of serum-supplemented cultures was as follows: phosphatidylcholine greater than phosphatidylinositol greater than sphingomyelin greater than phosphatidylethanolamine greater than phosphatidylserine. The distribution of glucose radiolabel into individual polar lipid classes of serum-free HTC cells was different from their serum-supplemented counterparts: sphingomyelin greater than phosphatidylcholine greater than phosphatidylinositol greater than phosphatidylethanolamine greater than phosphatidylserine. Glycerol from glyceride classes contained a higher percentage of radioactivity than the acyl moieties, with this percentage significantly elevated in serum-free cultures. The data indicate that, although glucose is a substrate for HTC cell lipids, other precursors present in the culture system also contribute to the lipid constituency of this hepatoma cell line.


A comparison of simplified methods for lipoprotein quantification using the analytic ultracentrifuge as a standard. Two simplified methods for quantitative lipoprotein analysis have been calibrated and compared with each other using analytic ultracentrifugation as a standard reference procedure. The first method was the Friedewald procedure and the second was an automated agarose gel electrophoresis system. Both procedures offer comparable quantitative lipoprotein analysis with potential for large scale screening purposes at low cost ($4.00-$5.00 per analysis). There were advantages and limitations to both procedures. The Friedwald procedure can be used on frozen sera but requires 3 ml sera. In contrast, the electrophoresis system must be used with fresh serum but requires only 50 mul serum and the electrophoretic slides may be quantitatively analyzed several years retrospectively.


The effect of a short term saturated fat diet on the apoprotein composition and radioiodination properties of rat very low density lipoproteins. The effect of a saturated fat diet on the apoprotein composition and radioiodination properties of plasma very low density lipoprotein (VLDL) was studied in rats. After feeding the diet for 10 days, the proportion of 125I attached to VLDL lipid decreased from 50% (control animals) to 8%, the remainder (92%) being bound to the apoprotein components. The decreased lipid labelling was associated with proportional changes in the fatty acid composition of serum and VLDL lipids, the most notable change being a reduction in linoleic acid (30-8%) content which occurred in all the major lipid classes of both serum and VLDL. Analysis of VLDL after radioiodination showed that most of the radioactivity incorporated into the lipid moiety was associated with phospholipid. The proportion of 125I bound to phospholipid decreased after feeding rats a saturated diet. The proportion of soluble (small molecular weight peptides and arginine rich peptide) to insoluble (B apoprotein) did not alter during the saturated fatty acid dietary regime and no differences in the distribution of soluble proteins were observed. It is concluded that feeding a saturated fat diet to rats for 10 days significantly improved 125I labelling of the apoprotein moiety while apparently not inducing changes in apoprotein composition.


Black-white similarities in cord blood lipids and lipoproteins. Cord blood lipoproteins were quantitated in 117 neonates (58 white, 50 black) to assess for potential early expression of racial lipid distinctions. In comparison of black and white neonates there were no differences in total cholesterol (TC), high-density lipoprotein cholesterol (C-HDL), low-density lipoprotein cholesterol (C-LDL), C-HDL/C-LDL, or C-HDL/TC. Cord blood triglycerides were slightly higher in black neonates (p = 0.02). Unlike certain adult black-white comparisons and within the limits of "genicity" as expressed by cord blood lipoproteins, there were no racial differences in C-HDL, C-LDL, and total cholesterol.


Urinary excretion of carnitine and serum concentrations of carnitine and lipids in patients with hypofunctional endocrine diseases: involvement of adrenocorticoid and thyroid hormones in ACTH-induced augmentation of carnitine and lipids metabolism. The promoting effect of ACTH on carnitine and lipid metabolism was studied in patients with various endocrine hypofunctions. The results were compared with those of normal subjects. In adrenocortical insufficiency, hypothyroidism and hypopituitarism urinary excretion of carnitine was significantly lower than in normal subjects. On intramuscular injection of synthetic beta1-24 ACTH-Z urninary excretion of carnitine in normal subjects increased sixfold on the day of the injection and returned to the pretreatment level on the third day. Serum concentrations of carnitine and FFA increase in parallel with carnitine excretion, while serum triglyceride was lowered in response to ACTH administration. These responses were totally lacking or substantially suppressed in patients with the above endocrine insufficiencies. In hypothyroid and hypopituitary patients substitution therapy restored the responses to ACTH in the same fashion as those in normal subjects. These findings suggest that the promoting effect of ACTH on carnitine and lipid metabolism requires the presence of intact adrenocortical and thyroid functions.


Mechanism of action of luteinizing hormone and follicle-stimulating hormone on the ovary in vitro. The mechanism of action of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) upon various cell types of the mammalian ovary is reviewed. Emphasis is placed upon in vitro studies using organ and cell culture as well as short-term incubations. FSH and LH actions upon the following ovarian functions are discussed: steroidogenesis and metabolism of the ovary as a whole and of the isolated follicle and its component cell types, the granulosa and thecal cells, as well as folliculogenesis and follicular growth, oocyte maturation, follicular rupture, and corpus luteum maintenance and steroidogenesis. The roles of gonadotropin receptors, AMP, prostaglandins, protein kinase, and protein synthesis in these LH and FSH actions are discussed. Intra-ovarian regulation of LH and FSH action is reviewed, including a discussion of the possible roles of follicular fluid inhibitors upon oocyte maturation and granulosa cell luteinization.


Neonatal familial hyperalphalipoproteinemia. A kindred with four-generation vertical transmission of familial hyperalphalipoproteinemia was ascertained by measurement of elevated levels of cord blood high-density lipoprotein cholesterol (C-HDL) in a neonatal propositus. Quantitation of cord blood C-HDL coupled with family studies and longitudinal follow-up allows the diagnosis of familial hyperalphalipoproteinemia in infancy.


Catecholamine inhibition of thyrotropin-induced secretion of thyroxine: mediation by an alpha-adrenergic receptor. Thyroxine secretion by mouse thyroid gland incubated in vitro was measured. Thyrotropin or dibutyryl cAMP increased thyroxine secretion several-fold. l-Epinephrine and l-norepinephrine strongly inhibited this stimulated release; l-isoproterenol was relatively ineffective. Phentolamine prevented the inhibition by catecholamines of thyroxine release; l-propranolol had no effect. These findings indicate that stimulation of alpha-adrenergic receptors opposes the action of thyrotropin in the regulation of thyroxine secretion.


Hepatocelluilar carcinoma: a possible complication of oral contraceptive steroids. A patient was found to have developed a primary hepatocellular carcinoma after taking an oral contraceptive steroid preparation for four years. After resection of the tumour over five years ago, and resumption of the birth control Pill, a recurrent hepatocellular caricnoma has developed. The risk of this complicationis extremely small and appears to be related to the duration of ingestion of the drug and to the amount of oestrogen present in the preparation used. The modes of presentation of these tumours and the methods of investigation are outlined, and the high incidence of spontaneous rupture causing life-threatening intraperitoneal haemorrhage is emphasized.


Hepatocellular carcinoma in association with androgen therapy. The case of a young man with hypogonadism, who was treated for five years withmethyltestosterone and who developed hepatocellular carcinoma, is discussed. He received a course of 5-fluorouracil via the hepatic artery. Complete regression has been maintained for four years. It is likely that the androgen therapy is implicated in the development of the liver tumour. Recent reports of untoward effects of steroid hormones are reviewed.


An extensive rotavirus outbreak in aboriginal infants in central Australia. Faecal specimens were obtained from 92 Aboriginal infants admitted to Alice Springs Hospital during an extensive outbreak of gastroenteritis in the surrounding area in July, 1976. Specimens were examined for potentially pathogenic viruses, bacteria and parasites. Rotavirus proved to be by far the most common potential pathogen, being detected in 54% of specimens.


[Diagnostic significance of identification of anti-entamoeba histolytica antibodies (author's transl)]. Examinations were performed with the indirect immunofluorescence test (IFAT) and the indirect haemagglutination test (IHAT). Negative titers (less than 1:5) do not exclude an infection with E. histolytica. Titers up to 1:80 in both reactions are found in patients with negative stool specimens or indicate an asymptomatic infection of the intestinal lumen, an incipient or past symptomatic amoebiasis. IFAT values of 1:160 together with a negative or positive IHAT are also suggestive of the two last-mentioned types of infection. These titers are also found in patients with gastrointestinal disorders of other orgin. IFAT values higher than 1:60 with a positive IHAT are probably due to an invasive amoebiasis. Titres are, however, only diagnostic pointers.


Development and applications of Bacillus subtilis test systems for mutagens, involving DNA-repair deficiency and suppressible auxotrophic mutations. A mutagen-tester of Bacillus subtilis was constructed and tested with known carcinogens. The parental strain HA101 of Okubo and Yanagida carrying suppressible nonsense mutations in his and met genes was transformed to carry an excision-repair deficiency mutation. The constructed strain TKJ5211 showed a 20--30-fold higher sensitivity for His+ reversion than the parental strain when treated with UV and UV-mimetic chemicals but unchanged mutation frequency with X-rays and methyl methanesulfonate. The tester strain was used in a spot test of 30 selected chemicals and also for testing with liver homogenate activation. The results showed an almost equivalent but somewhat broader detection spectrum than the Salmonella typhimurium TA100 system. Another test method used a pair of B. subtilis strains differing in their DNA-repair capacity, i.e. the most UV-sensitive mutant HJ-15 and a wild-type strain, to detect repair-dependent DNA damage produced by chemicals. Spores could be used in either test.


Repair of single-strand DNA breaks and recovery of chromosomal and chromatid aberrations after treatment of plant seeds with propyl methanesulfonate in vivo. Repair of single-strand breaks of DNA and simultaneous recovery of chromosomal aberrations were studied after treatment of barley seeds with the monofunctional alkylating chemical mutagen, propyl methanesulfonate in vivo. In soaked seeds the diminution of single-strand breaks of DNA induced by PMS was correlated with the decrease of chromosomal aberrations, whereas in dried seeds the repair of DNA breaks was depressed and, in accord with this, the frequency of chromosomal aberrations increased. The prolonged storage of seeds led to a more delayed repair of chromosomal aberrations in dry seeds and a more delayed accelerated repair in soaked seeds.


Nephritogenic glycoprotein. V. Immunochemical studies on nephritogenic activity of collagenase and pronase-digests prepared from various rat organs. A new immunochemical procedure was introduced to estimate the nephritogenic activity of collagenase and pronase digests of various rat organs. A glycoprotein isolated from collagenase digests of various rat organs showed the nephritogenic activity as well as the antigenic activity that induces nephrotoxic antibody, which were nearly identical to those of a glycoprotein isolated from trypsin digests of the rat organ concerned. A glycoprotein isolated from pronase digests of various rat organs was proved to have no antigenic activity that induces nephrotoxic antibody, but the existence of nephritogenic activity was proved in this glycoprotein. These results were supported firmly by the assay experiments for nephritogenicity. Ouchterlony gel diffusion test and the assay experiments demonstrated the existence of a common nephritogenic substance among the glycoproteins isolated from trypsin, collagenase and pronase digests of rat organs.


Interaction between parathyroid hormone and catecholamines on renal cortical cyclic AMP. Parathyroid hormone (PTH) and catecholamines (CA) increased cAMP levels in isolated tubules of rat renal cortex. A rise in cAMP could be detected 10 sec after the addition of agonists, and it reached the peak in 30-60 sec and then decreased despite the presence of active agonists. Direct relationships exist between theophylline concentrations and cAMP levels in response to agonists. Three- to fivefold rises in cAMP levels were observed with a maximal dose of PTH compared with that of CA, and there was no additive effect. Both PTH and CA stimulated renal gluconeogenesis; when added together with each maximal dose, no additive effect was observed. These results suggest the important role of phosphodiesterase to control tubular cAMP in response to hormonal stimuli, and there is a tubule cell group responding to both PTH and CA.


Adrenal cortical secretory responses to histamine and cyanide in dogs with hypothalamic lesions. Experiments were performed on dogs with hypothalamic lesions as well as on intact and hypophysectomized dogs. Adrenal venous blood samples were collected and analyzed for 17-hydroxycorticosteroids (17-OHCS). In intact dogs, a marked increase in adrenal 17-OHCS secretion was observed after an i.v. injection of histamine and following an i.v. infusion of cyanide. Adrenal 17-OHCS secretion in response to histamine was markedly reduced but not completely abolished by hypophysectomy and was partially impaired by anterior median eminence lesions and by posterior median eminence lesions, but was not significantly impaired by supramammillary lesions. Adrenal 17-OHCS secretion in response to cyanide was completely abolished by hypophysectomy, markedly reduced by posterior median eminence lesions and partially impaired by anterior median eminence lesions and by supramammillary lesions. Thus, a partial dissociation of the adrenocortical secretory responses to histamine and cyanide was observed in dogs with hypothalamic lesions, suggesting that the mechanism and the pathway to the anterior pituitary involved in the adrenocortical secretory response to histamine and cyanide are somewhat different.


Plasma glucocorticoid elevation and ultrastructural changes in the adenohypophysis of the male rat following prolonged exposure to stress. A morphological and ultrastructural study is described which indicates that cellular activity in the adenohypophysis correlates well with the circulating levels of corticosterone. Intense secretory activity is observed in all tropic cells of the adenohypophysis over 10 days; thereafter the cellular morphology shows a return to the control condition. There are, however, differences in the degree of adaptation between the different tropic cells. After its initial hyperactivity, corticotrope activity returned to a control level by 20 days. Thyrotrope activity was also found to adapt to control activity, but only after 40 days. Similar patterns were observed in the stomatotrope and gonadotrope, where the initial hypertrophy returned to control levels by 20 days; thereafter, however, an inhibition was observed. The luteotrope however, seems to be an exception in that its level of activity increased throughout the duration of the stress procedure.


Morphological aspects of the vessels of brain tumours. The vascular changes associated with cerebral tumours are demonstrated angiographically and histologically.


Lack of association between defective delinquents and antibody of herpesvirus hominis. Several groups have reported a relation between herpesvirus hominis infection and certain psychiatric disorders. We have investigated herpes antibody levels in chronic criminal offenders who were diagnosed as defective delinquents and in criminals who were not defective delinquents. We found no difference in the frequency of titers of herpesvirus type I or type 2 antibody in these groups.


[Observations on the therapy of endothoracic sarcoidosis with ACTH]. The Author on the basis of data obtained with ACTH treatment in endothoracic sarcoidosis concludes that the administration of corticotrophin is advisable mainly when an intolerability to corticosteroids is present or when corticosteroid therapy does not display any appreciable effect. When steroid treatment yields partially satisfactory results, ACTH administration can produce further improvements.


[Behavior of blood lipids and lipoproteins in myocardial infarct]. Blood lipids and lipoproteins were evaluated in male and female subjects of various ages with acute myocardial infarct. Values were within the limits of normal in less than 50%, mostly in the aged. II B (24%), IV (18%) and II A (11%) lipoprotidaemic pictures were the most common. It is felt that abnormal values are not necessarily part of the infarct picture; if present, they are mainly of the II B type.


Herpes simplex virus type 2 meningitis.

Herpes simplex virus type 2 meningitis. To date only 9 culture-proven cases of herpes simplex virus Type 2 meningitis have been recorded. Presented here is the 10th case with a review of the literature. Although generally a benign disease, its recognition in association with herpes progenitalis must be appreciated to avoid confusion in the treatment of this common veneral disease.


Herpes zoster ophthalmicus.

Herpes zoster ophthalmicus. Herpes zoster ophthalmicus is not an uncommon disease and is more prevalent among debilitated and seriously ill patients. It is caused by the same virus causing varicella. The exact trigger mechanism is unknown, as well as much of the pathogenesis. The disease is more uncommon among the elderly and usually runs a benign course. Approximately 50% of the patients develop ocular complications ,the most frequent of these being keratitis, iritis, secondary glaucoma and extraocular muscle involvement. The most striking pathologic features are the lymphocytic infiltration of the long ciliary nerves and the vasculitis of the vessels accompanying them. The most controversial aspect of the disease is that of treatment. Almost every therapeutic regimen has been attempted in a disease whose natural course is self-limited. The future will add more to our knowledge of the pathogenesis of the disease and shed more light on the efficacy of various treatments.


Aniridia-Wilms' tumour syndrome.

Aniridia-Wilms' tumour syndrome. The association of aniridia and Wilms' tumour constitutes a real syndrome, which is genetic. It may either be autosomal dominant or depend on a chromosomal deletion or also, according to Knudson's theory, be due to two mutational events, the initiating mutation being germinal and the promoting mutation being post-zygotical.


Prophylactic antibiotics in cataract surgery. The value of preoperative cultures, sensitivity tests and prophylactic preoperative antibiotic treatment was evaluated in a group of senile, uncomplicated lens extractions. Sulfacetamide or a mixture of chloramphenicol and polymixin were applied preoperatively for 1 week and an injection of penicillin and streptomycin was given postoperatively. A control group of 140 cases without treatment was also studied. No postoperative infections developed in either group. The number of pathogenic cultures was significantly smaller in the treated group as compared with the untreated one; the mixture of chloramphenicol and polymixin B sulfate was found to be superior to sulfacetamide, different clinical data was compared in the two groups. No postoperative infections developed in either group.


Study on the pathogenesis of bluetongue: replication of the virus in the organs of infected sheep. The pathogenesis of bluetongue infection was studied by the titration of the virus in tissue samples taken from sheep inoculated subcutaneously in the auricula of the ear with 76 TC ID50 of the plaque-purified type 10 bluetongue virus. Tissue samples were taken from individual animals killed at daily intervals over a period of 11 days. The mean incubation time was 6.9 days and the first clinical sign was pyrexia. On the 4th day, bluetongue virus was demonstrated in the lymph nodes of the cephalic area, tonsils and spleen; viraemia became demonstrable on the 6th day post-inoculation and typical macroscopic lesions due to the virus were first observed on the 8th day. It was concluded that, post-infection, the virus entered the regional lymph nodes. From there it was disseminated via the lymph and/or the blood stream to the lymphoid tissues in other parts of the body where further replication occurred. From these primary sites the virus was carried via the blood stream and infected the majority of tissues. Humoral antibody, as detected by immunofluorescence, did not appear to have a direct influence on the concentration of virus in solid tissues. Persistence of the virus in infected sheep was not demonstrated when tissues were taken 6, 8 and 16 weeks after infection.


Kinetics of the IgM and IgG immunological response to sheep erythrocytes and bluetongue virus in mice. The IgM and IgG response of mice to sheep erythrocytes (SRBC) and bluetongue virus (BTV) was determined by means of haemolytic plaque assays. Maximum primary IgM response to SRBC occurred after 4 days but declined rapidly to 4% of the maximum by Day 9. A lag period of about 2 days was observed in the appearance of IgG haemolytic plaque-forming cells (PFC) but they reached a maximum after 6-9 days. Secondary immunization resulted in the stimulation particularly of IgG PFC and from Day 6 onwards IgG predominated in the immunological response. The IgM response to BTV was remarkably similar to that observed when SRBC were used as antigen. IgG PFC, however, appeared within a day of the IgM, reaching a peak on Days 4-5. From then onwards, IgG PFC predominated in the response. At BTV concentrations of up to 10 mug per mouse, the virulent strain of BTV type 3 produced the weakest response. At higher antigen concentrations there was very little difference in the response to the serotypes tested, although the virulent strain of BTV type 4 tended to produce the strongest response.


The isolation and preliminary genetic classification of temperature-sensitive mutants of bluetongue virus. Temperature-sensitive mutants of bluetongue virus were isolated and classified in 6 genetic recombination groups. The frequency of recombination varied both within and between groups. The 4 mutagens used viz. nitrous acid, N-methyl-N-nitroso-N-nitroguanidine, proflavine and 5-fluoro-uracil were found to differ in their efficacy. The period of incubation required for maximum recombination was 48 h at 28 degrees C.


Glossopyrosis due to adenoid cystic carcinoma. Glossopyrosis and glossodynia may occur from local or systemic factors or from any irritation along the course of the lingual nerve. Although cylindroma is relatively rare, it must be considered along with other malignant lesions when neurologic symptoms of burning and pain of the tongue persist. Examination of the lesion is best carried out as multiple needle biopsies rather than as open biopsy, in order to avoid seeding of the skin and lymphatics of the neck. Recurrences are frequent and, because of the slow growth, there must be a long follow-up period. Metastases to the lung and brain, although late, occur in large numbers of patients. Best results are obtained by a combination of radiation therapy and operation because the infiltration is so extensive that operation alone may not eliminate the tumor completely.


[Infantile diarrhea and rotavirus (author's transl)]. The acute and convalescent sera of 328 children with acute enteritis were checked with the bovine diarrhea virus as antigen in complement fixation test. 13% had a seroconversion. Some cases were associated with skin rashes or signs of respiratory infection.


[Residual dyslipoproteinemias after ethanol-weaning in alcoholic hyperlipidemic patients (author's transl)]. Twenty-two hyperlipidemic and long-term alcoholic patients were chosen as subjects with a chronic alcohol-dependent hyperlipoproteinemia, since their lipidemia was getting normal after they stopped alochol ingestion. After ethanol-weaning of these subjects, we found quantitative abnormalities of lipidemia (hypocholesterolemia in 5 cases among 22) and qualitative abnormal electrophoretic patterns in all cases: There is always a VLDL stain, a reduction of the LDL and VLDL migration. These facts suggest that exists in these patients a constant abnormality either in the enzymatic, or in the lipoprotein structures, which is induced or revealed by alcohol ingestion.20


[Apolipoprotein abnormalities in low and very low density lipoproteins in a type V hyperlipemia (author's transl)]. Abnormalities in VLDL and LDL apolipoproteins have been observed in the serum of an 11 year old girl with a type V hyperlipoproteinemia (VLDL + chylomicrons). This was shown with polyacrylamide gel electrophoresis and also with two dimensional immunoelectrophoresis. The LDL contain peptides which are not found in normal apo LDL. In the VLDL, heterogeneity of the lipopeptides is more marked that in normal VLDL, and there is an abnormal distribution of the apolipoproteins C or D.


Deficient activity of hepatic pyruvate dehydrogenase and pyruvate carboxylase in Reye's syndrome. The activity of certain hepatic enzymes involved in carbohydrate metabolism was measured in postmortem samples from six cases of Reye's syndrome. The activities of the two exclusively extramitochondrial enzymes, glucose-6-phosphatase and fructose-1,6-diphosphatase, were all within the normal range. Activities of pyruvate carboxylase and pyruvate dehydrogenase, both of which are exclusively mitochondrial enzymes, were below levels, shown by control tissue in every case, the average being 21.7% of the lowest control value for pyruvate carboxylase and 11.6% of that for pyruvate dehydrogenase. Impaired pyruvate metabolism appears to be another feature in Reye's syndrome.


The clinical, biochemical, and familial presentation of type V hyperlipoproteinemia in childhood. Primary type V hyperlipoproteinemia was identified in two preadolescent children. The propositus (kindred N) was a 10-year-old girl with severely creamy plasma, lipemia retinalis, hypertriglyceridemia (triglyceridelevel, 6,800 mg/100 ml), and ypercholesterolemia (cholesterol level, 490 mg/100 ml). Her parents and an 8-year-old sister all had endogenous hypertriglyceridemia (type IV hyperlipoproteinemia). In kindred A, an 11-year-old boy had triglyceride levels as high as 1,100 mg/100 ml and recurrent abdominal pain. His father had type V hyperlipoproteinemia; his mother was normal. All three of his older teenage siblings had type IV hyperlipoproteinemia. The enzymatic activities of lipoprotein lipase (LPL), hepatic triglyceride lipase (HTL), and histaminase (H) were studied in postheparin plasma. The LPL level was low in the children and both parents in kindred N. LPL level in kindred A was normal, except for one child with type IV hyperlipoproteinemia. HTL level was normal to above normal in both kindreds. Most patients had a normal H level, but one parent (kindred N) had no preheparin H and very low levels of postheparin H. There was a strong correlation (r = 0.58, significant at less than 1% level) between release of LPL and H but not between HTL and H (r= 0.22). The mean (+/- 1 S.D.) levels of the enzymes were as follows: LPL, 2.8 +/- 0.7 micronmol/ml/hr in kindred N and 5.4 +/- 2.2 micronmol/ml/hr in kindred A; H, 13.4 +/- 6.8 units/ml in kindred N and 22.0 +/- 11.9 units/ml in kindred A; and HTL, 18.0 +/- 7.1 micronmol/ml/hr in kindred N and 14.9 +/- 6.3 micronmol/ml/hr in kindred A. The enzymatic activities of kindreds N and A were significantly different for LPL (P less than .001) and H (.025 less than P less than .05) but not for HTL. All but one child had at least one high insulin level, which was accompanied by hyperglycemia in two children. The hypertriglyceridemia in all but one child was ameliorated on therapeutic diets. These data suggest that the genetic basis of the hypertriglyceridemia in these two families is different and that hyperchylomicronemia in childhood is not confined to the rara type I hyperliporproteinemia.


Effects of preloading of stannous compounds on the distribution of 99mTc-pertechnetate. 99MTc-pertechnetate distribution studies were performed in rabbits and mice following pretreatment between 5--336 hours with various routinely used stannous complexes (HSA, MAA, GHT, DTPA, PYPs) containing different amounts of Sn++ (0.17--15.0  mu mg/kg). Beyond a concentration of 0.26 mu mg/kg of Sn++ an alteration in 99mTc-ertechnetate distribution was observed. The red blood cell was found to be the most prominent target. An in-vivo reduction of 99mTc-pertechnetate apparently occurred by the presence of stannous ion within the red blood cell. Preloading time period between 5--24 hours did not alter the uptake of RBC/plasma ratio. Beyond that period it decreased slowly and still persisted up to 2 weeks following pretreatment. RBC/plasma ratio of 99mTcO4-increased with increased Sn++ content of various commercially available pharmaceutical kits.


Teaching patients with peripheral vascular disease. The teaching of a patient with peripheral vascular disease offers many challenges to the nurse. Although content should be understood and utilized by the patient and significant others, of primary importance is the belief that the learner is the curriculum and that his perception of self and his problems is the focus of teaching. The learning needs generally should be considered as the needs of an adult individual, who has many past experiences and an interest in the present and future. This patient may experience pain from his disease and may require the assistance and support of others. If the purpose of treatment is to improve circulation and prevent trauma and infection, then patient instruction must be recognized as a vital tool in therapy. But the nurse cognizant of the progressiveness of peripheral vascular disease must capitalized on the teachable moments--when the patient is ready and the learning is needed.


Separation of subclasses of human serum high density lipoproteins by zonal ultracentrifugation. An improved one-step method for the preparative separation of three subfractions of high-density lipoproteins from normal human serum has been developed. It employs the method of rate zonal ultracentrifugation in a z-60 rotor using a discontinuous NaBr gradient in the density range of 1.0-1.4. The density gradients were monitored directly by a flow-through density meter allowing the direct read-out of the actual densities in the process of filling and emptying the rotor. The separation of the three density fractions from 5 to 15 ml serum was achieved during a single 12 hours run at 59.000 rpm. The three fractions showed characteristically different patterns on polyacrylamide gel electrophoresis and differences in their lipid and protein composition.


[Effect of cobamide coenzyme on the growth of several transplantable tumors in rats and on prolonging the life of rats with grafted Zajdel ascitic tumor]. The effect of the cobamide coenzyme (5,6-dimethylbenzimidazolyl-Co-5'-deoxyadenozylcobamide, DBC) on the growth of some forms of rat tumours and on the prolongation of survival of rats with implanted Zajdela ascites hepatoma was compared with that of cyanocobalamine (5,6-dimethylbenzimidazolyl-Co-cyanocobamide, CN-B12). The effect of DBC was shown to differ from that of CN-B12. DBC did not stimulate the growth of the investigated forms of tumours and prolonged survival of rats with implanted Zajdela ascites hepatoma. The rats displayed cancer resistance when DBC or CN-B12 were injected before implantation of tumours or Zajdela ascites hepatoma.


[Adrenocorticotropic function of the pituitary gland in endocrine diseases]. In examination of 123 patients with diabetes mellitus. Itsenko-Cushing disease, Addison's disease, thyrotoxicosis and adiposity there was revealed an increase in the content of the adrenocorticotropic hormone (ACTH) in the blood. Comparison of the ACTH and cortizol concentration in the blood permitted to suppose a different mechanism of the derangements revealed. An increase of the adrenocorticotropic function of the hypophysis in diabetes mellitus, Itsenko-Cushing disease and thyrotoxicosis was accompanied by a rise in the blood cortizol level. A fall of glucocorticoid function of the adrenal glands in Addison's disease and a relative hypocorticism in the patients with adiposity caused a compensatory intensification of the ACTH secretion.


[Effect of long-term administration of hydrocortisone on the activity of glucosephosphatase and fructosediphosphatase and the synthesis of RNA fractions in the liver of rats of different age]. Experiments were conducted on albino rats of 3 age groups (2--3, 8--12, and 24--28-month-old). A study was made of the influence of prolonged administration of hydrocortisone on the activity of glucoso-6-phosphatase, fructoso-1,6-diphosphatase and the synthesis of RNA fractionated by the thermic phenol method. An increase of the enzyme activity, of the specific radioactivity, of the relative specific radioactivity of the RNA fractions was less pronounced in old rats. A comparatively early occurrence of the phenomenon of "disruption" of the inductive mechanisms of the hormonal stimulation was observed in the same age group.


Cell-free translation of simian virus 40 early messenger RNA coding for viral T-antigen. Simian virus 40 (SV40) mRNA was isolated by hybridization of cytoplasmic RNA, from SV40-infected BS-C-1 monkey cells early in lytic infection, to SV40 DNA immobilized on Sepharose. The early viral mRNA, when added to a wheat-germ translation system, directed the synthesis of a unique class of products including a 90,000 molecular weight (Mr) polypeptide. It was found that this 90,000 Mr product as well as a prominent 17,000 Mr polypeptide could be specifically immunoprecipitated with hamster antiserum to SV40 T-antigen, but not with hamster control serum. Similar immunoprecipitation of extracts of SV40-infected cells with hamster anti-T serum yielded 90,000 Mr and 17,000 Mr polypeptides; these polypeptides were not found in immunoprecipitates of uninfected cell extracts. SV40 cRNA, prepared by asymmetric transcription of plaque-purified SV40 DNA, directed the cell-free synthesis of several products, including a 70,000 Mr polypeptide that could be specifically immunoprecipitated with anti-T serum. However, no T-antigen-related polypeptide was found in infected cells that corresponded in size to the major immunoprecipitated cRNA product.


Stoichiometry of GTP hydrolysis and tubulin polymerization. Microtubule formation from lamb brain tubulin isolated by affinity chromatography and freed of exchangeable nucleotide requires GTP for maximal rate and extent of polymerization. The nucleotide analogs guanylylmethylenediphosphate and guanylylimidodiphosphate fail to replace GTP; in addition, neither the presence of microtubule associated proteins nor 5 M glycerol relieves the GTP requirement. The relation of GTP concentration and microtubule formation shows an association constant K = 1 X 10(4) M-1; furthermore, GDP and guanylylimidodiphosphate are competitive inhibitors of GTP for polymerization. Using a rapid filter assay for microtubule formation that allows the quantitative analysis of early polymerization kinetics and correcting for GTP hydrolysis uncoupled from tubulin polymerization, a stoichiometry of two molecules of GTP hydrolyzed per mole of tubulin dimer incorporated into microtubules has been found.


[Stimulating effect of human chorionic gonadotropin on the activity of guanidinoacetate-N-methyltransferase of rat testes: role of cyclic AMP]. A study was made of the influence of human chorionic gonadotropin (HCGT) on the activity of guanidineacetate-N-methyltranspherase in human testes. Guanidineacetate-methyltranspherase proved to be stimulated in the testes of the sexually immature rats in vivo under the action of HCGT on the 19th day after birth and from the 24th to the 27th day after birth. The activity of the enzyme was also increased in adult rats weighing 35--40 g. The activity of guanidineacetate-methyltranspherase in the testes of rats was particularly sharply stimulated after a single administration of N6-O2'-dibutiryladenosine-3',5'-cyclophosphate against the background of a preliminary treatment of rats with HCGT for a period of 5 days. The enzyme stimulation induced by combined treatment of rats with HCGT and N6-O2'-dibutiryladenosine-3',5'-cyclophosphate was prevented by the administration of cycloheximide and actinomycine D. The activity of the enzyme decreased after the incubation of the rat testes tissue homogenate with HCGT.


[Role of hormones of neurohypophysis in ACTH and glucocorticoid secretion]. The absence of a strict relationship between the antidiuretic activity of the posterior lobe of the hypophysis and the ACTH-glucocorticoid secretion was shown in experiments on rats. A conclusion was drawn that the neurohypophysial hormones played no significant role in the realization of the reaction of the hypophysioadrenal system to various actions, and could influence only the spontaneous level of the ACTH-glucocorticoid secretion.


Heteroduplex analysis of avian RNA tumor viruses. Electron microscopic heteroduplex analysis of avian RNA tumor viruses has been undertaken by using 35S viral RNA and long, complementary DNA synthesized in vitro. In this initial study, heteroduplex molecules were formed between complementary DNA from Rous sarcoma virus [Prague B strain (Pr-B)] and RNAs from Pr-B and Rous sarcoma virus [Prague C strain (Pr-C)] and from their transformation defective (td) derivatives, td-Pr-B and td-Pr-C. In the case of heteroduplexes with the td viruses, a deletion loop was observed of the order of two kilobases in size and less than one kilobase from the 3' terminus of the RNA. This deletion probably spans part or all of the sequences of one or more genes in the nondefective sarcoma virus which are essential for cell transformation. The sizes and the positions of the deletions in the td-Pr-B and td-Pr-C viruses were slightly, but significantly, different. No nonhomology features were observed in the Pr-B-Pr-C hybrids, thus confirming the close genetic relatedness of the two viruses. All heteroduplexes contained a proportion of circular and dimer molecules. This observation is a direct demonstration that (-) strand DNA transcription begins at an internal position of the RNA genome, proceeds to the 5' end, reinitiates at the 3' end of the RNA, and copies the remainder of the viral genome. Other implications for models of RNA tumor virus replication are also developed from these data.


[Age-specific characteristics of the effect of various hormones on the lipolysis in the adipose tissue of rats under normal conditions and in starvation]. Experiments were conducted in vitro; a study was mode of the age peculiarities attending be invluence of adrenaline, ACTH, STH on the lypolysis in the adipose tissue of rats under normal conditions and in starvation. During the process of ageing there occurs no change in the adipose tissue sensitivity to the action of hormones stimulating the lipolysis, but the capacity to react to high hormone doses is decreased. Elevation of the basal lypolysis in the adipose tissue of young rats occurs during starvation; the lypolytic action of the hormones falls. Starvation fails to alter the basal lypolysis in the adipose tissue to react to the hormone action.