Development of neuromuscular transmission in a larval tunicate. 1. The time sequence of the development of acetylcholinesterase (AChE), acetylcholine (ACh) receptors and functional synapses on the embryonic muscle membrane in a tunicate larva (Halocynthia roretzi) was investigated in vivo.2. The fertilized tunicate egg was incubated in natural sea water at 9 degrees C. Sixty-eight hr after fertilization the free-swimming larva was hatched, which had six striated muscle fibres in the tail. The developmental stage of the embryo was indicated by the developmental hours after fertilization.3. The transmitter at the neuromuscular junction in the hatched larva is ACh. (i) Neuromuscular transmission was completely blocked by D-tubocurarine (1-5 x 10(-5)M). (ii) Eserine (5-10 x 10(-7)M) approximately doubled the time constant of the falling phase of miniature excitatory junctional currents (e.j.c.s). (iii) The reversal potential of the membrane response to iontophoretically applied ACh was -10 mV and similar to that of e.j.c.s. (iv) AChE was present on the muscle membrane surface.4. AChE activity became visible histochemically on the embryonic cell membrane in the presumptive muscle region as early as the late gastrula stage (27 hr after fertilization, 12 hr before the ACh response appeared).5. The response to iontophoretically applied ACh was present at 39 hr after fertilization but could not be evoked at 38 hr.6. Between 39 and 41 hr after fertilization, the ACh responses increased rapidly, then remained relatively unchanged until larval hatching.7. The stage of the initial appearance of the ACh response corresponded to the stage when the Ca current abruptly increased in the muscle membrane.8. The first sign of neuromuscular transmission was appearance of a giant excitatory junctional potential (e.j.p.) with uniform amplitude (about 15-20 mV) and slow time course (time constant of the falling phase of a giant e.j.c. was 23.4 +/- 6.9 msec, mean and S.D., at -60 mV and 11 degrees C).9. Within a few hours, these giant e.j.p.s disappeared and were successively replaced by medium-sized e.j.p.s and then e.j.p.s similar to those seen in hatched larvae (time constant of the falling phase of a miniature e.j.c. was 8.5 +/- 1.8 msec at -60 mV and 11 degrees C).


Modulation of cyclic nucleotide levels in peripheral nerve without effect on resting or compound action potentials. 1. Cyclic nucleotide levels and compound action potential magnitudes were measured in frog sciatic nerves following exposure to carbachol, isoprenaline and cyclic nucleotide related substances. 2. The resting cyclic AMP level was 2-4 p-mole/mg protein and the cyclic GMP level was 0-27 p-mole/mg protein in desheathed nerves. 3. Isoprenaline (100 micrometer) caused a twofold increase in cyclic AMP without affecting cyclic GMP levels. Carbachol (100 micrometer) caused a twofold increase in cyclic GMP without affecting cyclic AMP levels. 4. The phosphodiesterase inhibitor theophylline (5 mM) augmented both cyclic AMP and cyclic GMP. 5. The magnitude of the resting or compound action potential was not affected by isoprenaline, carbachol, or phosphodiesterase inhibitors. 6. The cyclic nucleotides and their butyryl derivatives did not affect the magnitude of the resting or compound action potential, either when applied alone or in the presence of a phosphodiesterase inhibitor. 7. In contrast to sympatic tissue we conclude that hormone mediated cyclic nucleotide metabolism in peripheral nerve is unrelated to control of axonal excitability.


Caffeine: its direct and indirect influence on reproduction. Caffeine constitutes the active drug principle in a number of commonly consumed beverages. Among North American adults, it holds the distinction as being probably the most widely utilized psychotropic drug. The compound is most often consumed in the form of coffee, which is derived from the bean of the tropical tree Coffea arabica or Coffea robusta. The unique pharmacologic and physiologic properties of caffeine have led to extensive research efforts, especially in the area of mutagenically and teratogenically mediated effects. In this regard, investigators have often overlooked the alterations in catecholamine, free fatty acid and cyclic nucleotide levels that the the presence of caffeine brings about. Such alterations are discussed in light of their effect on reproduction, and an attempt has been made to clarify the effects that the associated habits of alcohol and smoking have on the reproductive system.


Fulminant puerperal sepsis associated with aplastic anemia: the case for prophylactic antibiotic therapy. Fulminant puerperal sepsis due to Clostridium perfringens occurred in a primiparous 19-year-old woman who developed aplastic anemia during pregnancy. Although the risk of infectious complications among pregnant women with compromised host defenses has not been accurately determined, it appears to be increased, particularly in patients with granulocytopenia. It therefore seems reasonable to give antibiotic prophylaxis during the intrapartum period to immunosuppressed women who come to term. Consideration of the bacterial pathogens most likely to cause acute endometritis in the early puerperium has led us to recommend a short course (72 hours) of penicillin G, 4 million units q4h, and gentamicin, 1,5 mg/kg q8h, as an appropriate regimen for such patients.


3-Hydroxy-17-aralkylmorphinans as potential opiate receptor-site-directed alkylating agents. In an effort to develop opiate receptor-site-directed alkylating agents, a series of 3-hydroxy-17-aralkylmorphinans containing reactive groups was synthesized and tested for analgesic and opiate antagonist activity. Many of the target compounds exhibited the characteristics of agonists and, among this group, some were found to be active blockers of morphine analgesia. One of the more potent antagonists (41) was investigated further and it was found that while its action is specifically associated with opiate receptors, 41 could not be classified either as a competitive or noncompetitive antagonist in the classical sense. The duration of antagonist action in vivo of 41 and its in vitro receptor binding characteristics suggest that covalent association with opiate receptors is not not an important factor.


Structure and biological activity of (-)-[3H]dihydroalprenolol, a radioligand for studies of beta-adrenergic receptors. (-)-Alprenolol is a potent competitive beta-adrenergic antagonist. "(-)-[3H]Alprenolol", a radioactive form of this agent produced by catalytic reduction with tritium, has recently been used successfully as a radioligand for direct studies of beta-adrenergic receptors. In this communication it is documented that the compound formed by catalytic reduction of (-)-alprenolol with tritium gas is the saturated product (-)-[3H]dihydroalprenolol in which tritium is added across the double bond and exchanged into the adjacent benzylic position. No exchange into the aromatic ring was observed. These conclusions were substantiated by results obtained on hydrogenation and deuteration of (-)-alprenolol. The biological activity of (-)-[3H]dihydroalprenolol, dihydroalprenolol, and alprenolol was also shown to be identical as assessed by direct ligand binding and inhibition of catecholamine-stimulated adenylate cyclase.


Techniques for typing Herpesvirus hominis antibody: a comparison of inhibition of peroxidase-labelled antibody staining with inhibition of indirect haemagglutination and with microneutralisation. A new technique based upon the inhibition of the peroxidase-labelled antibody staining (PLAS) has been used to type Herpesvirus hominis (HVH) antibodies in four groups of human sera taken from patients with one or both types of HVH infection and in mixtures of different proportions of type 1 and type 2 antisera. The results were compared with those of the microneutralisation (MN) test and the indirect haemagglutination (IHA) inhibition test. The sensitivity and specificity of the three methods were identical for sera containing only one type of HVH antibody. The MN test was slightly more sensitive than the other tests for detecting small amounts of HVH-1 antibody mixed with large amounts of HVH-2 antibody. Nevertheless, the PLAS inhibition technique was far more rapid and it would seem a satisfactory alternative to the IHA inhibition test for HVH antibody typing.


Seroepidemiologic studies on the possible relationships of human and bovine leukemia: Brief communication. A seroepidemiologic study was conducted in an attempt to identify antibodies against the bovine leukemia virus (BLV) in people exposed to cattle with lymphosarcoma. Farm families, farm employees, and veterinarians in contact with cow herds having documented cases of lymphosarcoma were tested for precipitating antibodies to the BLV with the agar gel immunodiffusion test. The cattle also were tested serologically. Information was collected form the farm families regarding consumption of unpasteurized milk from their dairy herd. Twenty-one dairy herds with documented cases of lymphosarcoma were identified. A total of 846 cows from these herds wre bled, of which 33% were serologically positive. No positive sera were found in the 45 dairy farmers, family members, and farm employees associated with the herds with lymphosarcoma. Consumption of raw milk was reported by 77% of the farm group. In addition, 83 veterinarians, 30 leukemia patients, and 200 control human sera were tested and found negative for antibodies to the BLV.


Dissociation of antiviral and antitumor immunity in resistance to Marek's disease. Immunization of chickens either with gluteral-dehyde-inactivated chicken kidney cells infected with Marek's disease (MD) virus or with glutaraldehyde-inactivated cells of MD lymphoma-derived continuous lymphoblastoid cell lines protected against MD. The former type of immunity was associated with an immunologic suppression of virus replication and virus antigen production after challenge with virulent virus, but lymphocytes specifically cytotoxic to cells bearing MD tumor antigens were not detected. In the latter type of immunity, virus multiplication was not affected; some evidence of the stimulation of cell-mediated antitumor immunity was found. The results supported the view that immunity to MD may be directed against either virus-specific or tumor-specific antigens and that in natural resistance to MD both mechanisms may be operative.


Spontaneous regression of Friend virus-induced erythroleukemia. II. regression of Friend murine leukemia virus-induced lymphocytic leukemia. We characterized several aspects of spontaneous regression of lymphocytic leukemia in mice. The disease, induced by the helper murine leukemia virus (MuLV) component obtained from the regressing Friend virus complex (RFV), was characterized by spleen and lymph node enlargement, thymus involvement, and anemia. Leukemia regression occurred in about 25% of infected mice and resulted in the return of lymphoid organs to near-normal weight and normal histology and the recovery from anemia. A tenfold to 1,000-fold decrease in virus titer was seen in those mice in which leukemia regressed when compared to leukemic animals, although infectious virus was still recoverable from apparently normal spleens. The sera of mice in which leukemia regressed contained potent virus-neutralizing activity that was associated mainly with immunoglobulins. These studies firmly supported the evidence that the regressing phenotype of RFV was due to its helper MuLV component (MuLV-RF).


Hypocalcemia and nephrogenous cyclic AMP production in critically ill or injured patients. Critically ill or injured patients often have impaired cardiovascular function. Since low ionized calcium levels can cause such changes, serum calcium and urine calcium were measured in a prospective study involving 28 criticially ill or injured patients and 16 normal controls. Serum protein levels were also measured to calculate "corrected" total calcium levels. Ionized calcium levels are difficult to measure. Since ionic hypocalcemia is thought to increase the "nephrogenous production" of cyclic AMP, cyclic AMP levels were measured in the blood and urine of these patients and the "nephrogenous" cyclic AMP calculated from the creatinine clearance. The mean total serum calcium in these patients was 7.7 +/- 0.8 mg/dl (S.D.). This was significantly lower (p less than 0.001) than our controls (9.6 +/- 0.6). When corrected for hypoproteinemia, the mean serum calcium (8.7 +/- 0.8) was still significantly lower (p less than 0.005) than control (9.4 +/- 0.5). The mean urine calcium excretion in the patients (56 +/- 66 mg/100 ml G.F.R.) was lower, but not significantly so, than in the controls (84 +/- 44 mg/100 ml G.F.R.). The "apparent nephrogenous" cyclic AMP in the study group was 2,731 +/1 1,451 pm/ml/100 ml G.F.R. The nephrogenous cyclic AMP had a negative correlation (r =-0.45) with "corrected" total calcium levels. Thus "total," "corrected" total, and "ionized" calcium levels appear to be reduced in the majority of critically ill or injured patients studied. The clinical implications of these findings and the potential value of serial cyclic AMP determinations in blood and urine will be discussed.


Management of nasopharyngeal angiofibroma. Successful surgical treatment of nasopharyngeal angiofibroma requires complete pre-operative evaluation and careful choice of a suitable surgical approach. Experience with 12 patients presenting with this tumor demonstrates the value of polytomography and angiography in evaluation. The use of pre-operative estrogens and temporary vessel ligation has decreased blood loss. The surgical approach is modified depending upon tumor location. A tumor confined to the nasopharynx is removed by a transpalatal approach. Extension into the sinuses or orbit necessitates a sublabial incision or lateral rhinotomy. Pterygo-maxillary tumor may be exposed by extension of the sublabial incision posterior to the maxillary tuberosity. This allows one to push the tumor back into the nasopharynx. Intracranial extension is removed by a combined otolaryngologic-neurosurgical approach.


Pseudotypes of vesicular stomatitis virus with the envelope properties of mammalian and primate retroviruses. By employing improved techniques it has been possible to produce and characterize a representative spectrum of mammalian and primate retrovirus pseudotypes of vesicular stomatitis virus (VSV). Selection of appropriate cell lines for both the production and subsequent detection of the VSV pseudotypes has been the most important factor in permitting their demonstration. The host range for penetration of these retrovirus pseudotypes of VSV has been defined and found to differ from that reported for the replication of the corresponding retroviruses. Additionally, retroviruses having an identical host range for replication were distinguishable by differences in their host range for penetration, implying that restriction of replication may be occurring by different mechanisms. Studies of the plaque-forming efficiency of retrovirus pseudotypes of VSV in cell lines nonpermissive for replication of the corresponding retroviruses permitted a distinction to be made between the restriction of replication occurring as a consequence of postpenetration events and that occurring as a consequence of a block of penetration itself. The demonstration of primate retrovirus pseudotypes of VSV permits the use of VSV as a probe for the detection of this group of viruses.


Cell-free synthesis of herpes simplex virus-coded pyrimidine deoxyribonucleoside kinase enzyme. The incubation of a cell-free protein-synthesizing system prepared from rabbit reticulocytes with cytoplasmic RNA from herpes simplex virus (HSV)-infected cells resulted in increased thymidine kinase activity. This enzyme activity was specifically inhibited by anti-HSV antiserum and was relatively unaffected by TTP, an inhibitor of cellular thymidine kinases. Induction of the new activity was prevented by addition of inhibitors of eucaryotic protein synthesis, and no new activity was detected when RNA from cells infected with pyrimidine deoxyribonucleoside kinase-deficient mutants, instead of wild-type HSV, was added. An increased deoxycytidine kinase activity with similar properties to the HSV-specified enzyme activity was also present in cell-free systems incubated with RNA from HSV-infected cells. Phosphorylation of thymidine and deoxycytidine at 30 degrees C continued for longer than 11 h. The findings are consistent with the accurate synthesis in vitro of enzymically active HSV-specified pyrimidine deoxyribonucleoside kinase.


Effect of helper virus on the number of murine sarcoma virus DNA copies in infected mammalian cells. Cell lines of four mammalian species were each examined for the number of Moloney murine sarcoma virus (M-MSV) DNA copies in total cellular DNA after M-MSV transformation. Sarcoma-positive, leukemia-negative (S+L-) M-MSV-transformed cells were compared to M-MSV-transformed cells infected with a replicating leukemia virus. Both unfractionated M-MSV complementary DNA and complementary DNA representing the MSV-specific and the MSV-murine leukemia virus-common regions of the M-MSV genome were hybridized to total cellular DNA of various species. DNAs of mouse, cat, dog, and human S+L-cells contained from less than one to a few proviral M-MSV DNA copies per haploid genome. In contrast, helper virus-coinfected, M-MSV-producing cells of each species showed a 3- to 10-fold increase in M-MSV proviral DNA over that found in corresponding S+L- cells. MSV-specific and MSV-murine leukemia virus-common nucleotide sequences were each increased to a similar degree. A corresponding examination of cellular DNA of leukemia virus-infected normal or S+L- mammalian cells was performed to establish the resulting number of leukemia proviral DNA copies. The infection of normal or S+L- mammalian cells with several leukemia-type viruses that did not have nucleotide sequences closely related to the cell before infection resulted in the appearance of one to three corresponding leukemia proviral DNA copies.


Clonal transformation of adult human leukocytes by Epstein-Barr virus. We have developed a clonal transformation assay for Epstein-Barr virus which uses adult human leukocytes as target cells. The target cells were isolated from Epstein-Barr seronegative donors, and the same donor's cells could be studied repeatedly over long periods of time. When these cells were transformed by Epstein-Barr virus and had proliferated sufficiently to be studied, they had an average cloning efficiency of 3%. Assuming this average cloning efficiency obtains at the onset of transformation, we calculate that transformation by Epstein-Barr virus leads to immortalization maximally of about 1 in 30 of the adult peripheral leukocytes exposed to the virus. Studying the number of colonies transformed as a function of the amount of virus to which the cells are exposed indicates that a single DNA-containing virus particle is sufficient to transform a cell. All of the transformed clones studied harbored viral DNA. This technique will now permit, for the first time, our studying clonal variations in adult peripheral leukocytes transformed by Epstein-Barr virus as a function of input multiplicity of the virus and of the donor's immune status.


Relationship between Moloney murine leukemia and sarcoma virus RNAs: purification and hybridization map of complementary DNAs from defined regions of Moloney murine sarcoma virus 124. Complementary DNAs (cDNA's) specific for various regions of the Moloney murine sarcoma virus (MSV) 124 RNA genome were prepared by cross-hybridization techniques. A cDNA specific for the first 1,000 nucleotides adjacent to the RNA 3' end (cDNA 3') was prepared and shown to also be complementary to the 3'-terminal 1,000 nucleotides of a related Moloney murine leukemia virus (MLV) genome. A cDNA complementary to the "MSV-specific" portion of the MSV 124 genome was prepared. This cDNA was shown not to anneal to Moloney MLV RNA and to anneal to a portion of the viral RNA of about 1,500 to 1,800 nucleotides in length, located 1,000 nucleotides from the 3' end of MSV RNA. A cDNA common to the genome of MSV and MLV was also obtained and shown to anneal to the 5'-terminal two-thirds, as well as to the 3'-terminal 1,000 nucleotides, of the MSV RNA genome. This cDNA also annealed to the RNA from MLV and mainly to the 5'-terminal half of the MLV genome. It is concluded that the 6-kilobase Moloney MSV 124 RNA genome has a sequence arrangement that includes (i) a 3' portion of about 1,000 nucleotides, which is also present at the 3' terminus of MLV; (ii) an MSV-specific region, not shared with MLV, which extends between 1,000 and 2,500 nucleotides from the 3' terminus; and (iii) a second "common" region, again shared with MLV, which extends from 2,500 nucleotides to the 5' terminus. This second common region appears to be located in the 5' half of the 10-kilobase MLV genome as well. Experiments in which a large excess of cold MLV cDNA was annealed to (3)H-labeled polyadenylic acid-containing fragments of MSV RNA gave results consistent with this arrangement of the MSV genome.


Integration of proviral DNA in chicken cells infected with Schmidt-Ruppin Rous sarcoma virus is not enhanced by DNA repair. The effect DNA repair might have on the integration of exogenous proviral DNA into host cell DNA was investigated by comparing the efficiency of proviral DNA integration in normal chicken embryonic fibroblasts and in chicken embryonic fibroblasts treated with UV or 4-nitroquinoline-1-oxide. The cells were treated with UV or 4-nitroquinoline-1-oxide at various time intervals ranging from 6 h before to 24 h after infection with Schmidt-Ruppin strain A of Rous sarcoma virus. The chicken embryonic fibroblasts were subsequently cultured for 18 to 21 days to ensure maximal integration and elimination of nonintegrated exogenous proviral DNA before DNA was extracted. Integration of proviral DNA into the cellular genome was quantitated by hybridization of denatured cellular DNA on filters with an excess of (3)H-labeled 35S viral RNA. The copy number of the integrated proviruses in normal cells and in infected cells was also determined from the kinetics of liquid RNA-DNA hybridization in DNA excess. Both RNA excess and DNA excess methods of hybridization indicate that two to three copies of the endogenous provirus appear to be present per haploid normal chicken cell genome and that two to three copies of the provirus of Schmidt-Ruppin strain A of Rous sarcoma virus become integrated per haploid cell genome after infection. The copy number of viral genome equivalents integrated per cell treated with UV or 4-nitroquinoline-1-oxide at different time intervals before or after infection did not differ from the copy number in untreated but infected cells. This finding supports our previous report that the integration of oncornavirus proviral DNA is restricted to specific sites in the host cell DNA and suggests a specific mechanism for integration.


Endogenous RD-114 virus genome expression in malignant tissues of domestic cats. Endogenous xenotropic cat type C virus (RD-114)- and infectious feline leukemia virus (FeLV)-specific gene expressions were measured in spontaneous sarcomas carcinomas, and nonmalignant cat tissues by molecular hybridization for virus-specific RNA and competition radio-immunoassays for the major internal protein (p30) of these two viruses. The results indicate that RD-114 gene expression in sarcomas and carcinomas at both RNA and p30 levels is significantly higher than histologically normal tissues from cats free of cancer. In contrast, the levels of FeLV viral RNA and p30 are fount to be low or undetectable in the majority of these tumored and normal tissues examined. Whereas variability in the amounts of RD-114 OR FeLV RNA and p30 expressed is found in tissues from different cats, their expression is fairly uniform in multiple malignant tissues of the same cat. The finding of widespread occurrence of elevated RD-114 gene expression in sarcomas and carcinomas is consistent with our similar observation with natural lymphomas of domestic cats and suggests that expression of certain functions of this endogenous virus may be etiologically involved in the development of many different spontaneous neoplasms of cats.


Feline leukemia virus: biochemical and immunological characterization of gag gene-coded structural proteins. The major non-glycosylated structural proteins of feline leukemia virus have been isolated, and competition immunoassays have been developed for each. These proteins include the 27,000- to 30,000-molecular-weight major internal antigen designated p30, a 15,000-molecular-weight protein (p15), an acidic protein of 12,000 molecular weight (p12), and a highly basic 10,000-molecular-weight protein (p10). Immunologically and biochemically corresponding proteins of feline and murine leukemia viruses have been identified. and, on the basis of analogy to the known sequence of a prototype type C virus of mouse origin, the map order of the gag region of the feline type C viral genome has been tentatively deduced as NH2-p15-p12-p10-COOH. The demonstration of two feline leukemia virus gag gene-coded proteins, p15 and p12, expressed in the form of an uncleaved precursor in a mink cell line nonproductively transformed  by feline sarcoma virus provides indirect support for the proposed sequence.


HindIII and Sst I restriction sites mapped on rabbit poxvirus and vaccinia virus DNA. The DNAs of two closely related orthopoxviruses, rabbit poxvirus (RPV) and vaccinia virus (VV), were mapped by overlapping-fragment analysis using restriction endonucleases HindIII and Sst I. The exact arrangement of these fragments was accomplished by total digestion of isolated partial restriction products and by end-fragment determination. RPV and VV DNAs showed identical restriction patterns in an internal region comprising approximately 60% of the genome. The size, by electrophoretical analysis of the RPV DNA, was 118 X 10(6) daltons, some 6 X 10(6) daltons less than VV DNA. The two opposite terminal restriction fragments of RPV DNA cross-hybridized to each other.


Differential effect of arabinofuranosylthymine of the replication of human herpesviruses. The thymidine analog 1-beta-arabinofuranosylthymine (ara-T) has previously been found to selectively inhibit herpes simplex virus replication. At a relatively nontoxic conentration (50 microgram/ml), ara-T reduced herpes simplex virus yields by 4 to 5 log10. Ara-T was also effective in inhibiting the replication of varicellazoster virus (VZV) in vitro in human embryo fibroblasts, completely preventing VZV-specific cytopathic effects. The inhibition of VZV was reversible upon drug removal at 48 h after addition but was not reversible after 5 days of treatment. ara-T also reduced cell-free virus infectivity and the plaque-forming cell yield of VZV. Compared with the untreated controls, which demonstrated a 1-log10 increase over input plaque-forming cells at 24 h after infection, 50 microgram of ara-T per ml resulted in a 1-log10 decrease. In contrast to herpes simplex virus and VZV, cytomegalovirus replication was relatively resistant to ara-T. Neither cytopathic effects nor the incorporation of [3H]thymidine into acid-insoluble material in cytomegalovirus-infected cells was markedly affected. Analysis of the newly synthesized labeled DNA by CsCl buoyant density determinations indicated that the same relative proportions of cell and virus DNA were synthesized with or without added drug. Interpretation of these results with regard to virus-induced deoxypyrimidine kinase is discussed.


Viruses of Entamoeba histolytica. VII. Novel beaded virus. A third amoebal virus type was isolated from four different strains of Entamoeba histolytica. The virus was most frequently seen as a linear structure about 235 nm long and consisting of 14 beadlike structures about 19 nm in diameter. A "dimer" of twice the length and consisting of 28 beads was occationally encountered. The virus replicated in the nucleus, forming ordered arrays. Acridine orange staining of viral aggregates in infected nuclei suggested the presence of double-stranded nucleic acid.


Simian virus 40-specific ribosome-binding proteins induced by a nondefective adenovirus 2-simian virus 40 hybrid. We have studied the intracellular distribution of the two simian virus 40-specific proteins, with apparent molecular weights of 56,000 and 42,000, detectable in human KB cells infected by a nondefective adenovirus 2-simian virus 40 hybrid, Ad2+ND2. After a 20-min pulse of [35S]methionine, about two-thirds of the newly synthesized 56K protein and one-third of the 42K protein were found localized on the plasma membrane. The remainder of each protein was found in the cytoplasm, whereas the nuclear fraction was virtually free of either component. A significant portion of both proteins present in the cytoplasmic fraction was complexed to the 40S ribosomal subunits and was not removed by treatment with 0.5 M KCl. Moreover, the portion that was found free in the cytoplasm could bind preferentially and quantitatively to purified 40S ribosomes in vitro, leading us to propose that these simian virus 40 proteins may act as translational control elements in cells.


Evidence for recombination between N- and B-tropic murine leukemia viruses: analysis of three virion proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We have sodium dodecyl sulfate-polyacrylamide gel electrophoresis to analyze the virion proteins of an N- and a B-tropic C-type virus derived from the BALB/c mouse and 21 putative recombinants, designated XLP-N viruses, obtained from seven crosses between these N- and B-tropic viruses. All the XLP-N viruses are N-tropic but posses the XC plaque morphology of their B-tropic virus parent. Three virion proteins, p15, p30, and gp70, of the parental viruses each differ in electrophoretic mobility. Two recombinants were found that possess a p15 that comigrates with p15 of the B virus; 19 possess a p15 that comigrates with N virus p15. Sixteen recombinants possess a gp70 that migrates like the gp70 of the B virus: four have gp70 with an electrophoretic mobility like that of the N virus gp70. All 21 recombinants possess a p30 that comigrates with p30 of their N virus parent. Given the origin and phenotype of XLP-N viruses, these results would seem to provide good evidence that these viruses are recombinants.


In vitro synthesis of a unique RNA species by a T particle of vesicular stomatitis virus. A T particle of vesicular stomatitis virus, containing most of the L-gene region, has been isolated. In vitro, these T particles synthesize exclusively a small adenine-rich RNA that is complementary to the T-particle genome. Partial sequence analysis of this small RNA indicates that it is an RNA of unique sequence with a length of approximately 45 nucleotides.


Dissociation of polyoma virus by the chelation of calcium ions found associated with purified virions. Analysis of polyoma virions by X-ray fluorometry demonstrated that calcium (Ca2+) was associated with the purified virion. Treatment of purified virions with ethyleneglycol-bis-N,N'-tetraacetic acid (EGTA), which chelates Ca2+, and the reducing agent dithiothreitol caused the virions to dissociate. Electron microscopy revealed that the virions were dissociated to the capsomere level. Incubation of polyoma virions with 150 mM NaCl, 10 mM  EGTA, and 3 mM dithiothreitol was optimum for the dissociation reaction. The pH for the dissociation reaction ranged from 7.5 to 10.5. Cesium chloride density gradient centrifugation indicated that both EGTA and dithiothreitol were necessary for dissociation to occur; neither reagent alone dissociated the virus. The major protein product of the dissociated viral particles sedimented at 12S. Relationships between these experiments and the alkaline carbonate-bicarbonate dissociation of polyoma are discussed.


Synthesis of proteins and glycoproteins in cells infected with human cytomegalovirus. In cytomegalovirus-infected cells, the rate of protein synthesis was detected as two peaks. One occurred during the early phase of infection, 0 to 36 h postinfection, and the other occurred during the late phase, after the initiation of viral DNA synthesis. Double-isotopic-label difference analysis demonstrated that host and viral proteins were synthesized simultaneously during both phases. In the early phase, approximately 70 to 90% of the total proteins synthesized were host proteins, whereas approximately 10 to 30% were viral, even at a multiplicity of infection of 20 PFU/cell. Virus-related proteins or glycoproteins were referred to as infected-cell specific (ICS). Two ICS glycoproteins (gp145 and 100) were clearly detectable and were synthesized preferentially in the early phase of infection. Their synthesis was concomitant with stimulation of the protein synthesis rate. In the late phase of infection, approximately 50 to 60% of the total protein synthesis was viral and approximately 40 to 50% was host. The ICS proteins and glycoproteins detected during the late phase of infection were viral structural proteins. Infectious virus was not detectable until 48 to 72 h postinfection. An inhibitor of viral DNA synthesis, phosphonoacetic acid, prevented the appearance of the late-phase ICS proteins and glycoproteins, but there was little or no effect on early ICS glycoprotein synthesis. Radiolabeled ICS proteins and glycoproteins were identified by their relative rates of synthesis, by their different electrophoretic mobilities compared with those of host proteins and host glycoproteins, and by their similar electrophoretic mobilities compared to those of proteins and glycoproteins associated with virions and dense bodies of cytomegalovirus. Structural viral antigens in the infected-cell extracts were removed by immunoprecipitation, using F(ab')(2) fragments of cytomegalovirus-specific antibodies, and identified as described above. The last two criteria were used to identify viral structural ICS proteins and glycoproteins. Although approximately 35 structural proteins were found to be associated with purified virions and dense bodies, the continued synthesis of host cell proteins complicated their identification in infected cells. Nevertheless, seven of the nine structural glycoproteins were identified as ICS glycoproteins.


Structural components of the arenavirus Pichinde. Purified Pichinde virions grown in monolayers of BHK-21 cells were found to contain three major species of virion proteins as described previously (Ramos et al., J. Virol. 10:661-667, 1972). Two of the proteins were glycosylated (G1, molecular weight = 64,000; G2, molecular weight = 38,000) and were present in similar proportions on the outer surface of the virions. A third protein (N, molecular weight = 66,000) was not glycosylated and, in association with the viral RNA species, was the major protein component of the viral nucleocapsids. An estimate of the approximate number of molecules of these three major proteins per virion was made. Minor amounts of other proteins were also routinely observed in Pichinde virus preparations. None of the three major protein species were phosphorylated to any significant exten, nor did they contain sulfated components. Two virion RNA species (L and S), but no 18S rRNA species, were detected in Pichinde virus preparations obtained from infected BHK-21 cells.


Correlation of virus polypeptide structure with attenuation of poliovirus type 1. Several avirulent samples of poliovirus type 1 derived in the process attenuating the neurovirulent Mahoney strain show an altered virus capsid polypeptide, VP-1, on polyacrylamide gel electrophoresis of sodium dodecyl sulfate-disrupted virions.


Antibody-independent detection of virus-specific glycoprotein synthesis is oncornavirus-infected cells. A method is described for detecting the synthesis of avian and murine oncornavirus-specific glycoproteins without the use of antibodies raised against viral structural proteins. As applied to cells infected with avian tumor virus, the method served to resolve pr90, the precursor of the major envelope glycoprotein. A virus-specific glycoprotein of about 85,000 daltons, which has several properties expected to a precursor to gp69/71, was detected in cells infected with murine leukemia virus.


Analysis of polyoma virus DNA replicative intermediates by agarose gel electrophoresis. Agarose gel electrophoresis has been used to fractionate polyoma virus DNA replicative intermediates (RI) according to maturity. Approximate electrophoretic mobility versus maturity relationships were obtained for both intact (supercoiled) and nicked (relaxed) RI. There was considerable overlap between the supercoiled and relaxed RI populations after electrophoretic fractionation. Intact RI could be recovered from preparative agarose gels for further analysis by centrifugation, electron microscopy, re-electrophoresis, or nuclease digestion.


Genetics of resistance to phosphonoacetic acid in strain KOS of herpes simplex virus type 1. A DNA- temperature-sensitive mutant of herpes simplex virus type 1 exhibiting thermolabile DNA polymerase activity, tsD9, was shown to be resistant to phosphonoacetic acid (PAA) when plated at the permissive temperature. ts+ revertants of tsD9 were PAA sensitive and exhibited DNA polymerase activity intermediate between that of the wild-type virus and tsD9, indicating that both temperature sensitivity and sensitivity to PAA are controlled by the same gene. Since the position of tsD9 on the existing herpes simplex virus type 1 linkage map is known, the locus for PAA resistance--and therefore for the structural gene for viral DNA polymerase--has been identified.


Amebiasis. An increasing problem among homosexuals in New York City. During a five-year period at The New York Hospital, Entamoeba histolytica was identified in the stools of 20 men who had not traveled outside the New York area. All of the patients were found subsequently to homosexual. During this same period amebiasis was diagnosed in 30 men who had traveled; only two were homosexual. Of ten patients with E histolytica infection seen during the first year of this study, none were homosexual whereas eight of 11 patients in the fifth year were homosexual, suggesting a gradual increase during this period of this disease in the homosexual community.


Venereal transmission of enteric pathogens in male homosexuals. Two case reports. In two cases there was simultaneous or sequential occurrence of amebiasis, shigellosis, and giardiasis in male homosexuals. Enteric pathogens may, under the proper conditions, be venereally transmitted. In particular, the sexual practices of male homosexuals, most significantly, oral-anal contact, appear to provide the necesseary link for transmission. Evidence suggests that this is a growing problem.


Scintigraphic evaluation of suspected acute myocardial infarction. Ninety-one patients with chest pain suggestive of acute myocardial infarction were studied by static technetium Tc 99m stannous pyrophosphate scintigraphy and dynamic sodium pertechnetate Tc 99m cardioangiography. Twenty-three of 26 patients (88%) with acute transmural infarcts and 12 of 17 patients (71%) with nontransmural infarcts had abnormal static studies. In 45 patients with negative scintigrams, ECG or serum enzyme changes consistent with acute infarction failed to develop. Three false-positive static studies (6%) were recorded. Twenty of 43 (47%) patients with acute infarction had hemodynamic or structural abnormalities identified by cardioangiography. The dynamic study also proved helpful in localizing the site of infarction and in ruling out certain causes of false-positive static scintigrams.


Venereal causes of cytomegalovirus mononucleosis. Cytomegalovirus (CMV) mononucleosis developed in two men after sexual contact with a woman who had had a similar but medically unverified illness several months before. The CMV was cultured from the woman's urine and cervix. Evidence of recent infection with CMV was also found in a new sexual contact of one of the men. However, roommates of the infected patients remained well, and serologic evidence of CMV infection failed to develop. This small outbreak suggests that CMV may be acquired as a venereal infection that sometimes progresses to the mononucleosis syndrome.


[Types of hyperlipoproteinemia in young persons with ischemic heart diseases engaged in strenuous mental work]. The results subsequent to an examination of 250 males aged 30-47 suffering from ischemic heart disease and doing strenous mental work are presented. Two variants were used in phenotyping: the first was based on not too high criteria for the cholesterol and triglycerides level (250 and 140 mg%, respectively), the second one having as a groundwork upper limits of the normal value estimated by the authors (250 and 190 mg%, respectively). It was found that, depending upon the actual value of the upper normal limit for triglycerides hyperlipidemia is demonstrable in 64 or 56% of the males. Of most frequent occurrence were the II and IV types of hyperlipoproteinemia with the latter type prevailing in both variants. Most patients with ischemic heart disease demonstrated a fall of alpha-lipoproteins, as ascertained by their separation through disc-electrophoresis in polyacrylamide gel.


[Blood viscosity in ischemic heart disease with different types of hyperlipoproteinemia]. Fifty patients with chronic forms of ischaemic heart disease were examined, their blood viscosity being determined with the aid of a rotational viscosimeter, and their hematocrit and fibrinogen levels being measured. Some increase in blood viscosity was found in patients with ischaemic heart disease with a tension shift of 0.7 and 0.5 dyn/cm2. In analysing the interrelationship between the blood viscosity figures and the risk factors inherent in ischaemic heart disease it was found that smoking results in an increasing blood viscosity with low tension shifts. The highest blood viscosity with low tension shifts. The highest blood viscosity values were found in patients with IIb and IV types of hyperlipoproteinemia. A direct correlation was established between the level of triglycerides and blood viscosity with a tension shift of 0.7 dyn/cm2.


[Lipid content in the blood serum of students under nervous-emotional stress]. Some aspects of the lipids metabolism in the blood serum of students differing in their age, working and learning experience prior to entering the institute, followed up throughout the examination period, are presented. Subject to determination was concentration of total lipids and their principal fractions: phospholipids, free and estherified cholesterol, free fatty acids and triglycerides, accomplished by means of micro-thin layer chromatography. Investigations were conducted 5 weeks, 1-2 days before examination and 5 weeks thereafter. A study of the lipids metabolism under nervous-emotional stress in dynamics may serve as one of the methodological approaches enabling one to reveal in good time any changes in the lipids metabolism.


[Herpes simplex of the lid margin (author's transl)]. Isolated herpes simplex affecting the lid margin can be distinguished from herpes simplex involving the entire eyelid by characteristic features. The typical intermarginal form of herpes-simplex-blepharitis presents itself as an erosion of about 3 mm in length which may be well discerned in detail after fluorescein staining. The skin adjecent to the lesion may also exhibit superficial ulceration, possibly being covered by crusts. The affected portion of the lid shows localized and moderate swelling as well as pain upon pressure. Injection of the bulbar conjunctiva will characteristically be noted mainly in the area opposing the lid lesion. The regional lymph nodes are enlarged, and possibly tender upon palpation. Local medication with anti-viral ophthalmic ointment is effective within a few days.


[From acupuncture to iris diagnosis (author's transl)]. Scientific medicine is always encircled by a miscellany of medical fantasies, which come and go, and which offer a short-cut to diagnosis and treatment, and (very occasionally) both together. Oculodiagnosis, or Iridodiagnosis, has little place in honset ophthalmology, but still holds its devotees.


beta-adrenergic receptor coupled-adenylate cyclase of human fat cell ghosts. The effects of beta-adrenergic agonists such as isoproterenol, norepinephrine and epinephrine upon the adenylate cyclase activity of human fat cell ghosts were tested, each alone and in combination with the beta-blocking agent propranolol. Saturating concentrations of these agents showed a 2-6.5-fold increase of enzyme activity without addition of any artificial cofactors. Isoproterenol was more potent in stimulating the enzyme system than epinephrine and nor-epinephrine. Propranolol caused a dose-dependent rightward shift of the log-dose response curve of these beta-adrenergic agonists. The assay of human fat cell adenylate cyclase in vitro may provide a simple anc convenient assay system for the screening of beta-adrenergic drugs of potential therapeutic importance.


[Studies on the structure and metabolism of lipoprotein-X (LP-X), the abnormal plasmalipoprotein in cholestasis (author's transl)]. Recent results regarding the pathophysiology of hyperlipoproteinemia in cholestasis are reported. The isolation of an abnormal lipoprotein (Lipoprotein-X; LP-X) from the plasma of cholestatic patients was achieved by a combination of various physico-chemical techniques. Most of the plasmacholesterol in these patients is transported in form of this abnormal lipoprotein which is very rich in phospholipids and unesterfied cholesterol. LP-X represents a vesicle with a mean diameter of 700 A. Albumin takes part as a structural protein of the particle. Besides albumin, which seems to be located in an internal water compartment or to be covered with lipids. Apo-C and Apo-D are present as surface proteins. The lack of Apo-B in LP-X, the major apoprotein of normal low density lipoproteins, seems to be the reason for a disturbed endogenous feedback mechanism of hepatic cholesterol synthesis, which is strongly increased in cholestasis. The high specificity and power of the LP-X test as clinical-chemical parameter to demonstrate or exclude cholestasis finds its explanation in our knowledge about the origin of this abnormal lipoprotein in cholestasis. LP-X is formed when a lipoprotein normally excreted with the bile refluxes into the plasma stream to convert into LP-X. This formation depends only on certain physico-chemical requirements and is independent of an energy-providing or enzymatically regulated process. The biological halflife of LP-X is similar to that of normal plasmalipoproteins. However, enzymes of postheparin plasma as well as the lecithin: cholesterol acyltransferase do not seem to play a major role in the catabolism of lipoprotein-X, but only change some of the physicochemical characteristics of this vesicle.


Inhibition by somatostatin of adenosine-3',5'-monophosphate stimulated hepatic ribosomal protein synthesis. Somatostatin which inhibits the secretion of various pituitary and intestinal hormones has been suspected to exert these effects by inhibiting adenosine-3',5'-monophosphate accumulation in the respective endocrine gland. Our results, obtained by cell free protein synthesis and by sedimentation through sucrose gradients of ribosomes, prepared from rat liver after incubation with cyclic AMP and/or somatostatin also suggest this antagonism between somatostatin and cyclic AMP. In addition, they indicate that this antagonism is not restricted to endocrine tissues.


Nerve excitability: transition from descriptive phenomenology to chemical analysis of mechanisms. The electrical activity by which impulses are conducted along nerve and muscle fibers, is carried by Na-and K-ions moving across the excitable membranes due to increased ion permeability. -- A biochemical approach, initiated to elucidate the mechanism of the permeability changes, centered around the analysis of the properties and functions of the proteins, including enzymes, directly associated with the role of AcCh, in the excitable membrane. The results necessitated a fundamentally reformed concept of the role of AcCh. The four proteins specifically associated with the function of AcCh form a cycle which controls the rapid ion permeability changes of the membrane and permits the ion fluxes through dynamic gateways. A model has been elaborated that integrates biochemical, biophysical, and thermodynamic data; it permits the interpretation of many electrophysiological data in molecular terms. AcCh has basically the same function in conducting and synaptic parts of excitable membranes. The new concept has replaced the purely descriptive phenomenology of nerve impulse propagation by the analysis of the chemical mechanisms of nerve excitability and bioelectricity.


The procoagulant activity of human granulocytes, lymphocytes and monocytes stimulated by endotoxin. Coagulation and electron microscopic studies. Leukocytes from donor blood were separated by ficoll/Urovison density centrifugation into granulocytes, lymphocytes and monocytes. The cell fractions were suspended in a culture medium to which endotoxin of Salmonella enteritidis was added at a final concentration of 10 microgram/ml. Endotoxin-stimulated monocytes developed a very high tissue factor (thromboplastin) activity while in granulocytes an only negligible amount of tissue factor activity was detectable. The tissue factor activity measured in the preparation of the lymphocytes can be explained by contamination with monocytes. Eelectron microscopic studies showed the lysosomes of all monocytes to be enlarged and activated. Only a fraction of the granulocytes appeared degranulated with prominent vacuoles containing inclusion bodies. Possibly the high tissue factor activity of the monocytes triggers the development of the disseminated intravascular coagulation in the Shwartzman phenomenon.


The stereology of pulmonary alveolar macrophages after prolonged experimental exposure to tobacco smoke. Morphometric analysis was performed on alveolar macrophages obtained by bronchopulmonary lavage of rats exposed to tobacco smoke for periods up to 60 consecutive days. Smoke dosage levels were adjusted so as to be comparable to those of human smokers. Size distributions were obtained for the lavaged macrophage populations of treated and age-matched control animals following 30 and 60 days of smoke exposure. Electron microscopic stereology was used to quantitate alterations in the ultrastructure of the same macrophage preparations. Several significant changes were observed in macrophage morphology following exposure to tobacco smoke, with an estimated mean cell volume more than twice that of controls following 60 days of smoking. After 30 and 60 days of exposure there was a 10- and 16-fold increase, respectively, in the volume density of cytoplasmic lipid inclusions. The surface to volume ratio of the cells and the lysosomal volume density were reduced in macrophages from smoke-exposed animals. A possible relationship between the incidence of lysosomes and the accumulation of cytoplasmic lipid in treated cells is discussed.


Cell shape and hexose transport in normal and virus-transformed cells in culture. The rate of hexose transport was compared in normal and virus-transformed cells on a monolayer and in suspension. It was shown that: 1) Both trypsin-removed cells and those suspended for an additional day in methyl cellulose had decreased rates of transport and lower available water space when compared with cells on a monolayer. Thus, cell shape affects the overall rate of hexose transport, especially at higher sugar concentrations. 2) Even in suspension, the initial transport rates remained higher in transformed cells with reference to normal cells. Scanning electron micrographs of normal and transformed chick cells revealed morphological differences only in the flat state. This indicates that the increased rate of hexose transport after transformation is not due to a difference in the shape of these cells on a monolayer.


Isolation of the alanine carrier from the membranes of a thermophilic bacterium and its reconstitution into vesicles capable of transport. A carrier protein mediating alanine transport was purified from the membranes of the thermophilic bacterium PS3, by ion exchange chromatography in the presence of both Triton X-100 and urea. The alanine carrier was recovered in the nonadsorbed fraction from either DEAE- or CM-cellulose columns, suggesting that its isoelectric point was in the neutral pH region. The final preparation contained virtually no electron transfer components, ATPase, or NADH dehydrogenase. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate revealed that the final preparation consisted of two major protein components with molecular weights of 36,000 and 9,400. Active transport of alanine after incorporation of the alanine carrier into reconstituted proteoliposomes was driven not only by an artificial membrane potential generated by potassium ion diffusion via valinomycin but also by mitochondrial cytochrome oxidase incorporated into the same liposomes and supplemented with both cytochrome c and ascorbic acid. The membrane-integrated portion (TFo) of the ATPase complex uncoupled alanine transport by conducting protons across the membrane.


Mutations affecting the binding, internalization, and lysosomal hydrolysis of low density lipoprotein in cultured human fibroblasts, lymphocytes, and aortic smooth muscle cells. Studies comparing the metabolism of low density lipoprotein (LDL) in normal cells and in cells cultured from patients with homozygous familial hypercholesterolemia have disclosed the existence of a receptor for plasma LDL. This receptor has been identified on the surface of human fibroblasts, lymphocytes, and aortic smooth muscle cells. An extension of these studies to cell strains derived from patients with other single gene defects in cholesterol metabolism has provided additional insight into the normal mechanisms by which cells regulate their cholesterol content and how alterations in these genetic control mechanisms may predispose to atherosclerosis in man.


Chlorpromazine inhibits cholera toxin-induced intestinal hypersecretion. The effect of chlorpromazine on cholera-toxin-induced intestinal fluid secretion (experimental cholera) has been tested in vivo in the mouse. It is shown that intramuscular injection of chlorpromazine 1 hr prior to toxin challenge effectively inhibits the net secretory response of the intestine. It is suggested that this effect might be due to inhibition of adenylate cyclase since chlorpromazine inhibits the increase of cyclic AMP concentration in vitro in mouse thymocytes incubated with cholera toxin.


Influence of dibutyryl cyclic AMP on the level of catecholamines and 5-hydroxytryptamine in discrete areas of rat brain. Dibutyrul cyclic AMP (dbcAMP) was injected intracerebroventricularly into rats in doses of 50, 100 or 200 microgram. Noradrenaline (NA), dopamine (DA), 5-hydroxytryptamine (5-HT) or 5-hydroxy indoleacetic acid (5-HIAA) concentrations were measured spectrophotofluorimetrically in discrete brain areas: cortex, hypothalamus, striatum, hippocampus with nucleus amygdala and nucleus accumbens (limbic system) and pons with medulla oblongata. DbcAMP (in each dosage administered) decreased, NA content of cortex and of pons with medulla oblongata. NA content in the hypothalamus and striatum was decreased only after the largest dose of dbcAMP. In the limbic system NA concentration was diminished after a dose of 100 or 200 microgram of dbcAMP. There was a decrease in DA level measured in the striatum alone after a dose of 100 microgram of dbcAMP. 5-HT concentration was increased in the striatum and the pons with medulla oblongata after each dose level of dbcAMP; in the cortex after a dose of 50 and 100 microgram; and in the hypothalamus after a dose of 200 microgram. 5-HIAA level was increased in the striatum, limbic system, pons with medulla oblongata after all doses of dbcAMP. We conclude that dbcAMP affects presynaptic central catecholaminergic and serotoninergic neurons in an opposite, manner, although discrete brain areas have a different susceptibility to dbcAMP action. These findings provide some explanation for the behavioural effects of dbcAMP.


[Islet cell tumors. Diagnostic and operative aspects (author's transl)]. Islet cell tumors have been described the past 25 years in frequent series. Multiple symptoms lead often to prolongation of disease and misinterpretation of diagnosis. Because of its rarity most authors report only few cases. We had the chance to observe 24 patients of which four had carcinoma within 12 years. Symptoms, clinic, different surgical procedures are described. Clinical difference seems to exists between benigne and malignant islet cell tumors.


Malignant fibrous histiocytoma of the head and neck. Review of the literature reveals of 29 cases of fibrous histiocytoma of the deep structures of the head and neck. Three additional cases are reported: a multicentric case involving the submandibular gland with metastases to the lungs; and intraosseous tumor of the mandible locally invasive; and a submental tumor which rapidly recurred after simple excision. A metastatic potential of 22% is established. The historm, nomenclature, histopathology and clinical features are reviewed. The value of electron microscopy and tissue culture in arriving at the current diagnosis are stressed. Wide excision is the treatment of choice.


Chronic otitis media and hearing loss in the Eskimo population of Canada. Chronic otitis media and hearing loss is a major health problem today in the Canadian Eskimo. The subject is reviewed and the findings of projects and a survey that have been carried out recently are discussed. A modality of treatment is proposed which is based upon the findings of the projects, the survey and other information that is available at the present time.


Levels of cyclic 3'-5' -adenosine monophosphate (cAMP) in maintenance cultures of rat hepatocytes in response to insulin and glucagon. Under conditions of short-term stimulation of lipogenesis by insulin in maintenance cultures of hepatocytes from starved rats, basal levels of cyclic 3'-5' -adenosine monophosphate (cAMP) are invariant. Glucagon signals increased levels of cAMP and severely diminishes lipogenesis. Insulin partially counteracts both glucagon effects.


[Effect of chronic action of cadmium sulfate on various biochemical indices of blood serum and on the histological picture of rat liver and kidneys]. Male rats weighing 180-220 g were given CdSO4, 0,4 mg/kg body weight subcutaneously once a week for 6-12 months. The animals were killed after 6, 9, 12 months and following blood serum levels were determined: total protein, albumin, globulin, GPT, GOT, Al.P. and urea. The tissue tissue samples from liver and kidneys were examined histologically (acid and alkaline phosphatase). After 9 months, the difference between values of biochemical indices in the exposed and control groups was statistically significant. It has been found that the observed biochemical indices show correlation with the extent of morphological changes in liver and kidneys. These degenerative changes became more intense in the liver than in the kidneys.


[Level of blood serum lipids in rats treated with detergents]. Male Wistar rats were treated intraperitoneally once per week for 12 weeks with following detergents: Olbrotol-18 (nonionic detergent), a product of etheric condensation of 18 moles of ethylene oxide to 1 mole of the mixture of olein alcohol and cetyl alkohol in ratio 1:1, in a dose of 10 mg/kg; SBO (anionic detergent), sodium 2-ethylhexylsulfosuccinate, in a dose of 10 mg/kg and Sterinol (cationic detergent), benzalkonium bromide, in a dose of 0.6 mg/kg. The control rats were injected with 0.9% saline solution. The content of total cholesterol, beta-lipoproteins and total lipids in serum were estimated. The increase of total cholesterol and the decrease of beta-lipoproteins content in serum of rats after all used detergents were observed as compared with the control. The increase of total lipid content only after long-term treatment with Olbrotol-18 was found. It is concluded that long term intraperitoneal treatment with detergents changes similarly the contents of total cholesterol and of beta-lipoproteins in blood serum of rats.


[Behavior of various biochemical indices of liver efficiency and of selected blood serum electrolytes in workers exposed to beryllium]. In 30 workers being occupationally exposed to beryllium, examinations of biochemical indicators of liver efficiency were carried out: activity of alanine and asparagine aminotraspherase and basic phosphatases, cholinesterase, content of total protein and its fraction. Levels of electrolytes were determined: calcium, potassium, phosphorus, and magnesium. The above examinations were also carried out on 30 persons who have no contact with beryllium. The obtained results were subjected to statistical analysis. In 10 persons from the group exposed to beryllium one found lowering of the level of magnesium in blood serum, whereas in the control group the level of this electrolyte was correct in all persons. As to the results on the level of magnesium, in both groups high statistical significance was found (p less than 0,01). A dependence was found between the amount of workers and the lowered level of magnesium in blood serum and the duration of occupational exposure to beryllium. The comparison of the remaining results of examinations of both groups did not reveal any statistically significant differences or the differences were at the point of statistical significance.


[Specific surface factor of dust particles and preliminary selection of silica dusts]. The results of particles analysis of three kinds of silica dust by means of miscroscopy and sedimentation in Andreasen's pipette served as a base for the determination of factor phi specific for particle surface. This factor is expressed as a ratio of the mean projected diameter (dp) to the aerodynamic diameter (d). The formula of the factor is: (formula: see text). The value of factor phi for tested silica dusts ranges from 2.0 to 2.6 (mean value--2.3), while the variability coefficient is 13.5%. Taking into account the value of factor phi determined by experiment and the value of dispersion of silica dust particles, the expected permeability of the selector (elutriator) and the deposition of dust in lungs were calculated.


Evidence that a separate particle containing B-apoprotein is present in high-density lipoproteins from perfused rat liver. High-density lipoproteins (HDL) (1.075 less than d less than 1.175) from perfusates of rat liver, unlike those of blood plasma, contain protein with the properties of B-apolipoprotein. This protein remains near the origin upon electrophoresis in polyacrylamide gels containing sodium dodecyl sulfate, has beta electrophoretic mobility in agarose gel, is insoluble in tetramethylurea, and precipitates with antisera to the B-apoprotein isolated from low-density lipoprotein. B-apolipoprotein in HDL from perfusates binds to concanavalin-A Sepharose and can thus be separated from the characteristic HDL, the chemical and physical properties of which are otherwise preserved. These observations suggest that in addition to the discoidal lipoproteins, another particle that contains B-apoprotein exists in HDL of perfusates.


[Mechanism of action of glucose on L-asparaginase synthesis by Escherichia coli bacteria]. The synthesis of L-asparaginase in Escherichia coli W and E. coli K-12 was almost completely supressed if glucose was added at a concentration of 0.5 per cent to a growth medium. The level of L-asparaginase synthesis decreased by ca. 75 per cent as a result of cyamutations when the bacteria could not produce cyclo-3',5'-AMP (cAMP). Apparently, a decrease in the intracellular content of cAMP caused by glucose could not be the only factor inhibiting L-asparaginase synthesis. Lactate was found to stimulate L-asparaginase synthesis. Glucose caused the catabolite repression and catabolite inhibition of the components of a system involved in lactate transport. The inhibition of L-asparaginase synthesis by glucose seems to be due, at least partly, to the fact that it prevents the assimilation of lactate by the cells, as well as the utilization of some other compounds which stimulate synthesis of this enzyme.


The effect of exogenous oestrogens on depression in menopausal women. A study of the effects of conjugated oestrogens (Premarin) upon the depression scores of depressed and non-depressed menopausal women was carried out. Depression levels were obtained for both groups before and after four weeks of hormone replacement therapy. The Beck Depression Inventory (BDI) was used to assess depression levels, while a semistructured interview provided information related to the physiological, psychological and sociocultural stresses affecting individual women. Depression scores of the depressed group did not improve within the given treatment period, in contrast with improved scores for the non-depressed subjects who reported increased feelings of well-being following hormone replacement therapy.


Solid phase indirect radioimmunoassay: standardization and applications in viral serology. In solid phase indirect radioimmunoassay (IRIA) antiviral immunoglobulins (Ig) bind specifically to the viral antigen fixed onto wells of microtiter plates. Radioactively labeled (125I) anti-Ig can be used to detect these antiviral antibodies. The sensitivity of the IRIA depends on the amount of antigen in the microtiter wells and the concentration of the 125I-labeled anti-Ig used. To standardize the assay, a method of antigen titration in the IRIA was devised, using parainfluenza type 1 viruses as antigens. The IRIA provides a means to titrate viral antigens with different biologic activities and antibodies against them. A binding inhibition test (BIT) based on IRIA allows the antigenic analysis of different virus strains, as is demonstrated for two closely related parainfluenza type 1 virus strains.


The site of synthesis and functions of acute phase plasma proteins: close relationship with the reticulo-endothelial system. The evidence that the acute phase glycoproteins of plasma are synthesized by the liver parenchymal cells is critically examined, and is found to be inconclusive. Some evidence is cited which favors the reticulo-endothelial system (RES) in general, and Kupffer cells in particular, as the site of synthesis of these proteins: 1. The entire RES contains non-glycogenic periodic acid Schiff-positive substances. 2. The diseases which affect glycoprotein levels are also known to affect the function of the RES. 3. When the animal is subjected to stress, the RES function is initially depressed and then stimulated. A similar biphasic behavior is shown by plasma glycoprotein levels. 4. Adrenal cortico-steriods are the major regulators of the RES function and of the synthesis of acute phase proteins. Moreover, both are stimulated at low concentrations, and depressed at high concentrations of the hormone. Some of the glycoproteins of the acute phase (prothrombin, the third component of complement, haptoglobin, transferrin and ceruloplasmin) have defense-related functions. The others seem to participate in phenomena like detoxification, promotion of phagocytosis, wound healing, prevention of tissue injury by lysosomal enzymes, prevention of trauma and recovery from inflammation. It is proposed that the acute phase proteins, together with antibodies, form major components of the definse system, and the RES attempts to deal with injury by mobilization of increased amounts of these substances.


Roles of prostaglandins suggested by the prostaglandin agonist/antagonist actions of local anaesthetic, anti-arrhythmic, anti-malarial, tricyclic anti-depressant and methyl xanthine compounds. Effects on membranes and on nucleic acid function. We have recently found that local anaesthetic, anti-malarial, anti-arrhythmic, tricyclic antidepressant and methyl xanthine compounds behave as prostaglandin antagonists at drug concentrations readily attainable in human body fluids. We have also found that various hormones, including prolactin, vasopressin and angiotensin when present in physiological concentrations can be potent stimulators of prostaglandin synthesis. Cortisol has no effect on basal prostaglandin systhesis but in physiological concentrations is able to reverse the effects of other hormones. The drugs are thought to act mainly by interfering with either calcium interactions with membranes, or with cyclic nucleotide synthesis or degradation or with nucleic acid function. We propose that prostaglandins play crucial roles at all three points. The hypothesis leads to new approaches to many aspects of cell regulation, to the functions of many systems, especially the neuromuscular, cardiovascular, renal and respiratory systems. It also has profound implications for drug design and evaluation and for the understanding of many clinical disorders.


Xanthine dehydrogenase from Drosophila melanogaster: a comparison of the kinetic parameters of the pure enzyme from two wild-type isoalleles differing at a putative regulatory site. Xanthine dehydrogenase (XDH) from Drosophila melanogaster has been purified to homogeneity by immunoaffinity chromatography, and its kinetic parameters determined. Drosophila XDH exhibits ordered binding for substrate and NAD+, analogous to the corresponding enzymes from vertebrate sources. The wild-type enzyme exhibits a Km for xanthine of 2.4 X 10(-5) M, and for NAD+ of 4.0 X 10(-5) M. XDH purified from a genetic variant exhibiting elevated levels of enzyme activity has similar kinetic constants. The results provide further evidence that the site of variation in the latter strain results in higher steady state numbers of XDH molecules per fly.


Methionine biosynthesis in Saccharomyces cerevisiae. II. Gene-enzyme relationships in the sulfate assimilation pathway. In Saccharomyces cerevisiae, the products of eleven different genes are needed for a functional sulfate assimilation pathway. Only five enzymatic steps are known in this pathway. The study of the gene-enzyme relationships has shown that the enzymes catalysing two of these steps are probably heteropolymeric. Moreover, mutations in three unlinked genes lead to multiple enzymatic losses. Different hypotheses are made to account for these results.


Characterization of a regulatory mutant of fructose 1,6-bisphosphatase in Saccharomyces carlsbergensis. The fdp mutation has been localized on the genome of Saccharomyces carlsbergensis, on chromosome II, between lys2 and tyr1, at a man distance of 31 centimorgan from lys2. Since the fdp mutant does not grow on glucose, fructose, mannose and sucrose, hexose transport and a number of enzymes of carbon metabolism were tested, but no significant differences could be found between the wild type and the mutant. Only the regulatory properties of glycogen synthetase are changed in the mutant, but it is doubtful whether this can explain its phenotype. The disorganization of carbon metabolism of the mutant upon addition of glucose to the medium was analyzed in more detail. The most prominent feature observed until now is the accumulation of free glucose and hexose phosphates in the cell. This result indicates that somehow the feedback control between hexose transport and metabolism is impaired. Hexose phosphates are known to be toxic to many cells, including yeast. Therefore, accumulation of hexose phosphates in the presence of glucose in the medium, can explain the absence of growth on this carbon source.


Mutants of Saccharomyces cerevisiae resistant to carbon catabolite repression. Mutants with defective carbon catabolite repression have been isolated in the yeast Saccharomyces cerevisiae using a selective procedure. This was based on the fact that invertase is a glucose repressible cell wall enzyme which slowly hydrolyses raffinose to yield fructose and that the inhibitory effects of 2-deoxyglucose can be counteracted by fructose. Repressed cells were plated on a raffinose--2-doexyglucose medium and the resistant cells growing up into colonies were tested for glucose non-repressible invertase and maltase. The yield of regulatory mutants was very high. All were equally derepressed for invertase and maltase, no mutants were obtained with only non-repressible invertase synthesis which was the selected function. A total of 61 mutants isolated in different strains were allele tested and could be attributed to three genes. They were all recessive. Mutants in one gene had reduced hexokinase activities, the other class, located in a centromere linked gene, had elevated hexokinase levels and was inhibited by maltose. Mutants in a third gene were isolated on a 2-deoxyglucose galactose medium and had normal hexokinase levels. A partial derepression was observed for malate dehydrogenase in all mutants. Isocitrate lyase, however, was still fully repressible.


Isolation and characterization of yeast mutants defective in intermediary carbon metabolism and in carbon catabolite derepression. Yeast mutants deficient in the constitutive ADHI (adc 1) were used for the isolation of mutants with deficiencies of the intermediary carbon metabolism, and of mutants defective in carbon catabolite derepression. Mutants were recognized by their inability to grow on YEP-glycerol and/or on ethanol synthetic complete medium. They were either defective in isocitrate lyase (ic11), succinate dehydrogenase (sdh1), or malate dehydrogenase (mdh1, mdh2), mdh-mutants could not uniformely be appointed to one of the known MDH isozymes. Homozygous mdh and sdh1 diploids are unable to sporulate. Three gene loci could be identified by mutants pleiotropically defective in many or all of the enzymes tested In ccr 1 mutants, derepression of isocitrate lyase, fructose-1,6-diphosphatase, ADHII and possibly of the cytoplasmic MDH is prevented, whereas the mitochondrial TCA-cycle enzymes, succinate dehydrogenase and malate dehydrogenase, are not significantly affected. CCR2 and CCR3 have quite similar action spectra. Both genes are obviously necessary for derepression of all enzymes tested. It could be shown that ccr1, ccr2 and ccr3 mutants are not respiratory deficient.


Endonuclease for apurinic sites in yeast. Comparison of the enzyme activity in the wild type and in rad mutants of Saccharomyces cerevisiae to MNS. It was found that yeast cells contain an endonuclease specific for apurinic sites in DNA which has no effect on DNA with normal strands or on strands with alkylated sites. The enzyme activity was studied in the RAD strain and in rad 6, rad 18-2 and rad 21 mutants, all very sensitive to MMS, as compared to the wild type. The level of endonuclease activity does not differ much between the tested strains, regardless of their differences in susceptibility to MMS. The enzyme activity is not induced by pretreatment of the cells with this mutagen.


Protein inhibitors of phosphorylase phosphatase and cyclic AMP-dependent protein kinase from rabbit skeleta muscle. A heat-and acid-stable protein inhibitor of phosphorylase phosphatase is present in a highly purified preparation of protein inhibitor of cyclic AMP-dependent protein kinase from rabbit skeletal muscle. Although these two inhibitors have strikingly similar properties to each other, such as sensitivity to trypsin and behavior on gel permeation chromatography, they can be separated by polyacrylamide disc gel electrophoresis. This indicates that the phosphatase-inhibitory and kinase-inhibitory activities reside with different protein species. The inhibition of both the enzymes is not altered by incubating the inhibitor preparation with a general phosphoprotein phosphatase, with phosvitin kinase, or with cyclic AMP-dependent protein kinase. Inhibition of phosphorylase phosphatase is of a non-competitive type supporting the idea that the phosphatase inhibitor is not an alternative substrate for the enzyme. Inhibition of phosphatase activity is selective in that it does no occur when phosphorylated histone or phosphorylated protamine are used as substrates.


Inhibition of rabbit skeletal muscle phosphorylase phosphatase by spermine. The effect of three naturally occurring polyamines (putrescine, spermidine, and spermine) on the activity of rabbit skeletal muscle phosphorylase phosphatase was investigated. Only spermine significantly inhibited the enzyme. The mode of inhibition (ki value of 0.3 mM) of the phosphatase by spermine appears to be different from that caused by divalent metal ions or by other organic cations, such as arginine and lysine esters, since it is noncompetitive with respect to the substrate, phosphorylase a.


Studies on the adrenal cortex of hypophysectomized rats: a model for abnormal cellular atrophy and death. Biochemical and ultrastructural studies indicate that the atrophy of adrenal cortex in hypoyhysectomized rats involves the following changes: (1) One to two days after hypophysectomy, there is loss of "template activity" resulting from cumulative DNA-damage and heterochromatinization. In vivo ACTH-administration led to recuperation of these cells, indicating damage during hypophysectomized state to be reversible. (2) If the duration of hypophysectomy is prolonged, some of the cells become irreversibly damaged and can no longer recuperate after in vivo ACTH administration. (3) The period of most rapid cell death is from the third to seventh day after hypophysectomy. The cause of cell death is probably due to membrane damage in the absence of protein synthesis, leading to lysis of the cells. Lysozomes and macrophages are apparently not involved.


[Electromyography as a diagnostic aid (author's transl)]. Electromyography reflects the anatomical situation in the muscle parenchyma. Changes in the anatomical structure are present if the normal potential form breaks up into small amplitude or even polyphasic potentials (myopathy type) or increases into high amplitude, broad and coarsely fragmented potentials (chronic neuropathy with regeneration). The caliber and type of muscle fibers are expressed in the speed of conduction. Spontaneous activity suggests an increased excitability of the muscel fibers. Disturbances of neuromuscular transmission are detectable by direct stimulation of the muscle. Analysis of nerve conductivity permits the demonstration of diffuse (polyneuropathy type) or circumscribed (entrapment neuropathy type) nerve affections. The recording of the parameters mentioned is the domain of EMG.


Activation of a procarcinogen to a mutagen by cell-free extracts of anaerobic bacteria. The Salmonella mutagenicity assay can be coupled to cell-free preparations derived from anaerobic bacteria (Clostridium perfringens and Bacteroides fragilis) to activate a procarcinogen to a mutagen. This activity is destroyed by heating and by digestion with pronase and it is sensitive to oxygen. These findings indicate that the Salmonella mutagenicity assay can be adapted to the study of the role of anaerobes in the activation of carcinogens.


The cytomegalic inclusion cell and disease; a hypothesis. Infections due to human cytomegalovirus are common in the general population, disease on the other hand is rare, being usually associated with the presence of cytomegalic inclusion bearing cells in the tissues. It can be inferred that in healthy individuals latently infected with this agent, such cells are promptly destroyed by the policing immune system and the efficiency of this immune surveillance is due to the high degree of antigenic disparity of cytomegalic inclusion bearing and normal cells. However, any impairment of the immune defence may render the host unable to deal with the inclusion bearing cells, which are then believed to induce at least some of the severe clinical symptoms of cytomegalic inclusion disease.


Mutagenic effectiveness and efficiency of sodium azide versus ethyl methanesulfonate in maize: induction of somatic mutations at the yg2 locus by treatment of seeds differing in metabolic state and cell population. This study was conducted to compare the effectiveness and efficiency of sodium azide tNaN3) and ethyl methanesulfonate (EMS) for inducing somatic mutations at the yg2 locus in maize seeds of two different metabolic states and cell populations. Dormant or presoaked (72 h at 20 degrees C) seeds heterozygous for yg2 locus were treated with different concentrations of either EMS or NaN3. The cell populations with respect to the percentage of cells in G1, S, G2, and M were also determined for seeds of the two metabolic states. Dormant seeds possessed a higher percentage of cells in G1 and the presoaked seeds a higher percentage of cells in S, G2, and M. The frequency of yg2 sectors in leaves 4 and 5 increased with increasing concentration of both mutagens in both dormant and presoaked seeds. Both mutagens were more effective and efficient in the presoaked seeds. NaN3 was more effective than EMS in terms of number of sectors induced per unit of dose. However, EMS was more efficient as determined by sectors induced per unit of seedling injury and clearly had the ability to induce much higher sector frequencies (more than 10 times greater) than NaN3. The low ability of NaN3 (compared to EMS) to induce mutant sectors may be related to the cells not being treated at the optimum time during the cell cycle, but it is more likely due to its low effectiveness for inducing chromosome aberrations.


Immunological selection of tumour cells which have lost SV40 antigen expression. In an already tumorigenic spontaneously transformed mouse cell, after further transformation by SV40, the virus-specific antigenic function becomes dominant. By transplantation into syngeneic mice SV40 antigen negative revertant tumour cells can be selected out.


Stimulation of calcium uptake into aortic microsomes by cyclic AMP and cyclic AMP-dependent protein kinase. Enhancement of calcium uptake into rabbit aortic microsomes was seen at a cyclic AMP concentration of 10(-6) M in the presence of cyclic AMP-dependent protein kinase (ATP: protein phosphotransferase, EC 2.7.1.37). Protein kinase alone also increased calcium uptake, but cyclic AMP alone was without effect. The results suggest that stimulation of calcium sequestration may be the mechanism of cyclic AMP involvement in vascular smooth muscle relaxation.


Increased level of cAMP in the rat intestinal mucosa caused by sodium lauryl sulphate. The level of cyclic AMP in the jejunal mucosa from tied loops of anaesthetized rats was found to be significantly increased (27-50%) when sodium lauryl sulphate (SLS) was added to the loop fluid (2-27 mM). Imidazole (25 mM) did not significantly alter the resting level of cyclic AMP, but reduced the increase caused by SLS (17 mM). Theophylline (25 mM) significantly increased the intestinal level of cyclic AMP, and potentiated the increase caused by SLS. Ouabain (2.5 mM) did not alter the level of cyclic AMP in the presence or in the absence of SLS. The results of previous experiments on the increases in intestinal absorption caused by SLS or by dibutyryl cyclic AMP (Briseid et al., 1974, 1976) are discussed in light of the present data. It is concluded that the SLS-effect on absorption can only partly by ascribed to its effect on the intestinal level of cyclic AMP.


The induction of tumors in Rana temporaria with nitrosamines. Dimethylnitrosamine (DMNA) and diethylnitrosamine (DENA) dissolved in tank water in a dose of 5 and 50 ppm, resp. induced tumors of liver (hepatocellular cancers and adenomas), and of hematopoietic system (hemocytoblastosis). DMNA induced neoplasms in 19 of 43 frogs (44.2%); the average latent period being 18.2 weeks, while DENA induced tumors in 41 of 94 animals (43.6%) within a mean latent period 19.7 weeks. Results of this investigation show the possibility to use anuran amphibians in shorttermed experiments in the study of nitrosocompounds carcinogenicity as well as a biological indicator for environmental pollution.


About the influence of halogenated pyrimidines on the in vivo-oncogenesis. Reported are in vivo trials on Friend virus-leukemia (FVL), on methylcholanthren-induced fibrosarcomas of the mouse, and on amelanotic melanoma No. 3 of the Syrian hamster after treatment with halogenated pyrimidines, 5'-bromodeoxyuridine (BUDR) and 5'-iododeoxyuridine (IUDR) as to provoke genuine oncorna viruses. Till now, no prove exists that in tissue culture propagated C-type particles are really oncogenic. From the results demonstrated here the conclusion can be drawn that the chosen in vivo-conditions neither oncogens, possibly located in the cells, will be finished to complete oncogenic viruses, nor a spread and multiplication of dormant viruses will take place due to a provocation treatment of the hosts with halogenated pyrimidines. It seems, therefore, that the requirements to activate oncogens in vitro are absolutely different to those needed under in vivo conditions.


[Computerized axial tomography in the diagnosis of multiple brain tumors-correlation with angiography and nuclear scanning (author's transl)]. Results of Computerized Axial Tomography with and without intravenous contrast (60% Conray) in 24 patients with multiple brain tumors were analyzed. Of the 24 patients, 19 had metastatic brain tumors, 4 multiple gliomas and 1 patient had multiple meningiomas. Data obtained by C.T. scan were compared to results obtained by angiography and nuclear scanning. In the diagnosis of multiple metastatic brain tumors. C.T. scans were superior to the other two procedures. however, C.T. scanning detected 4 lesions in the case of multiple meningiomas, while angiography disclosed 11 tumors in the same patient. In one patient having multiple gliomas, one (vascular) lesion only was demonstrated by angiography. Many other evaluations of Computerized Axial Tomography have been reported and the advent of this unique non-invasive radiologic technique has resulted in marked changes in the neuro-radiological diagnostic approach to brain tumors. However, it is important to recognize the weak as well as the strong points of each diagnostic procedure including the C.T. scan. At present, in our opinion, even with a positive demonstration of a brain tumor by C.T.scanning, cerebal angiography should be carried out prior to surgical intervention.


Comparative in vitro studies on corticotropin releasing activity of vasopressin and hypothalamic median eminence extract. The in vitro corticotropic releasing effects of vasopressin (VP) and hypothalamic median eminence (HME) extract were compared as a function of their concentration and preincubation and incubation times. Whereas HME extract augmented the ACTH secretion from non-preincubated adenohypophyses, VP released corticotropin from the pituitaries only after a 2 h preincubation period. The disappearance of endogenous VP during the preincubation time rendered the gland responsive. The maximal stimulation of ACTH secretion by VP was markedly less than that induced by HME extract. The results suggest the presence of VP receptor sites in the anterior pituitary (AP) which are probably different from the receptor sites of the hypothalamic corticotropin releasing factor (CRF).


EEG and task performance after ACTH4-10 in man. Effects of the heptapeptide ACTH4-10 (Org OI-63) on EEG, some memory tests, and behavior were examined in 12 normal male volunteers under 30 years of age. An intravenous dose of 60 mg was compared to placebo in a Latin square design. EEG was recorded for 2 h after administration of ACTH4-10 or placebo and was quantified by power spectral density analysis. Drug differences were tested by analyses of variance and covariance. No statistical drug effect was seen on the EEG or behavioral measures. Of the psychological tests, only the digit span test showed a decrease in number of errors with ACTH4-10 (p less than or equal to o.05). These results are consistent with previous studies and suggest that an intravenous bolus of ACTH4-10 has a limited effect on the brain function tested.


ACTH4-10: cognitive and behavioral effects in hyperactive, learning-disabled children. A single dose of 30 mg i.m. ACTH4-10 or placebo was given to a sample of 20 hyperactive, learning-disabled children. No significant drug effects were obtained on measures of visual and auditory memory, new learning, impulsivity, attention, perceptual motor skills, anxiety, or behavior during testing. There was a slight increase in pulse rate for drug compared with the placebo group. These findings are in keepeing witth other recent reports of limited or insignificant cognitive effects in adults of a single dose of this peptide.


Neuroleptics reduce spinal fluid cyclic AMP in schizophrenic patient. Cyclic AMP in the cerebrospinal fluid (CSF) was determined in a group of 10 schizophrenic patients before neuroleptic drug treatment and after a mean of 8 week's antipsychotic drug therapy. For 8 patients with marked to moderate treatment response a significant (p less than 0.01) decline in CSF cyclic AMP was observed. This result is consistent with the theory that blockade of postsynaptic dopamine receptors is a major mechanism of the antipsychotic action of neuroleptic drugs.


ACTH 4-10: a study of toxicological and behavioral effects in an aging sample. Placebo and ACTH 4-10 (in ascending dosages from 15 to 60 mg) were administered subcutaneously to normal aging subjects (mean, 65.6 years). Measurements were obtained for EEG, EKG, SMA-12, urine, and blood pressure. Behavioral tasks measuring reaction time (RT), short-term memory, perceptual speed, and motor speed were administered. There were no pre- to post-injection changes in SMA-12, urine, EEG, and EKG. There was an improvement in RT time that was dose related. These data indicate that ACTH 4-10 is (1) safe to administer to an aging population and (2) combined with data on young adults suggest that it may act on the attentional/arousal processes. Results suggest that ACTH 4-10 may have stimulant-like properties on behavior without effecting the CNS and cardiovascular system.


Effect of rubidium, lithium and cesium on brain ATPase and protein kinases. The authors studied the effect of rubidium, lithium and cesium on the ATPase system and c-AMP protein kinase in brain. They demonstrated that rubidium could replace potassium in the Na+K-ATPase system, whereas lithium and cesium had no effect on this enzyme activity in the absence of potassium. K+-dependent ATPase was activated by even low rubidium concentrations; lithium and cesium inhibited it. None of three (rubidium, lithium and cesium) affected c-AMP protein kinase.


EEG sleep diagnosis of medical disease in depression. The diagnosis of medical disease in the context of a depressive syndrome which may mimic medical illness has traditionally relied on a combination of exhaustive medical screening and neuropsychological testing. When 10 patients with primary depression were compared to 10 patients whose depression occurred in the context of concurrent medical disease, a single EEG sleep variable, total phasic REM activity (RA), correctly identified 95% of all 20 patients as either primary depressives or medical patients with depression. Conventional psychiatric assessment and neuropsychological testing were significantly less powerful discriminators among this sample.


Computed tomography of intra- and juxtasellar lesions. A radiological study of 108 cases. During the first two years with the 160 X 160 matrix EMI scanner at Rigshospitalet, Copenhagen 108 consecutive patients referred with the suspicion of intra- or juxtasellar tumor were subjected to 166 computed tomography (CT) examinations. The X-ray attenuation and contrast enhancement patterns of the various lesions were analyzed. In general, it was difficult to correlate these parameters with the histopathological features. Arachnoid cysts, however, had typical low preinjection attenuation and no contrast enhancement. Chromophobe and eosinophilic pituitary adenomas rarely contained calcium and only in minute amounts, hardly visible on the polaroid pictures. Craniopharyngiomas and low grade suprasellar gliomas frequently contained large calcifications. Grade I gliomas, when located in the optic nerves or hypothalamus, showed significantly higher contrast enhancement than elsewhere in the brain. Three purely intrasellar adenomas were demonstrated with CT only. The diagnostic accuracy of CT was compared to that of carotid angiography, PEG and plain skull films in the lesions verified by initial operation (n = 32). CT gave the highest accuracy of the four methods, but the accuracy of CT differed statistically only from that of carotid angiography.


Application of the radioreceptor assay for human chorionic gonadotropin in pregnancy testing  and management of trophoblastic disease. A commercially prepared radioreceptor assay (RRA) for human chorionic gonadotropin (hCG) has been evaluated as a pregnancy test and in a quantitative assay to follow patients with hydatidiform mole. The RRA demonstrated almost 100% agreement in comparison with radioimmunoassay (RIA) and urinary hCG tests. In the quantitative assay, a limiting reliable concentration of 70 mIU/ml of hCG in serum could be obtained. Extremely good correlation was achieved between the RRA and RIA test for hCG in 2 patients with hydatidiform mole over a span of 3 months of followup after evacuation of the mole. The usefulness of the RRA as a replacement of RIA tests for hCG is discussed.


Inhibition of growth of Morris hepatomas 7777 and 7800 by corn oil. Intraperitoneal injection of trace amounts of corn oil prior to and following the injection of 40-50 mg of tissue from hepatoma 7777 or 7800 into the thigh of adult male Buffalo rats resulted in a marked decrease in the growth rate of both tumors. Exhaustive extraction of the corn oil with water indicated that the active component was not water soluble. Similar injections of safflower oil or isotonic saline had no effect on tumor growth rate. Analysis of the tissue phospholipid fatty acids revealed that the injected corn oil caused no change in the esterified fatty acids in this lipid fraction.


The use of hybrid cells in immunotherapy. HARRIS et. al. isolated somatic hybrid cells (A9/SEWA) between polyoma-induced tumor and mouse fibroblast cell lines. Although these hybrid cells were no longer tumorigenic, we found that their immunogenicity was conserved. It therefore seemed to us that these antigenic, non transplantable, living cells would be an ideal tool for immunotherapy experiments. In our first experiments we assessed the immunoprotection afforded by A9/SEWA cells in both the SEWA tumor/A.SW mouse and C3HPy/C3H mouse systems. However, the efficiency of immunization with hybrid cells is dependent on the stability of the cells, especially in respect to the expression of the TATA. So we also tried to evaluate the immunogenicity as a function of the number of subcultures undergone by the hybrid line. In both systems, the immunogenicity was very good in the early subcultures but, in the C3HPy tumor/C3H mouse system, a loss of immunogenicity was observed as the number of subcultures increased. Thus any clinical application or immunization by hybrid cells would necessitate the verification of the presence of the tumor-associated antigens at each subculture. We are at present experimenting with various in vitro techniques for detection of the expression of these antigens.


Atypical fibrous histiocytoma of the orbit: an electron-microscopic study. A young woman developed progressive proptosis of her right eye, accompanied by retinal striae, exposure keratopathy and a severe decrease in visual acuity, reflecting optic neuropathy. An atypical fibrous histiocytoma was found within the orbit. She received orbital exenteration and irradiation. 2 years later she is free of clinical recurrence.


Oral granular-cell tumors. Report of twenty-five cases with electron microscopy. Twenty-five cases of granular-cell tumor of the oral cavity are reported. The clinical findings are similar to previously published data. Of five cases studied by electron microscopy, three were processed directly from fresh surgical specimens and two cases were processed following initial formalin fixation. The granules for the most part are morphologically compatible with lysosomes except for a single variety containing viruslike particles. There is increasing evidence in support of a neural origin, but the indications are that more than one cell type may be the progenitor of granular cells of the granular cell tumor.


An ultrastructural study of eosinophilic granuloma: the Langerhans cell--its role in histogenesis and diagnosis. An electron microscopic study of a case of eosinophilic granuloma of the mandible was performed. The fine structural similarity between the "histiocytic" cells of the lesion and epidermal Langerhans cells is discussed with regard to current theories and information on the origin of Langerhans cells and their potential role in the histogenesis of eosinophilic granuloma and other variants of histiocytosis X.


Histogenesis of salivary gland neoplasms. In an attempt to explain the histogenesis of salivary gland tumors, a bicellular theory of origin has been presented. This theory is supported by indirect evidence from light and electron microscopy. Two cells, the excretory duct reserve cell and the intercalated duct reserve cell, are presented as the hypothetical cells of origin for salivary gland neoplasms. It is argued that the excretory duct reserve cell gives rise to squamous cell carcinomas and mucoepidermoid carcinomas, and that the intercalated duct reserve cell gives rise to all others. It It is also shown that myoepithelial cells are responsible in part for the wide histologic variation of these neoplasms.


[Lipoprotein-x determination in children beyond an age of 6 months (author's transl)]. In premature and newborn infants an abnormal lipoprotein (LP-X) was almost regularly detectable by examination up to the 24th week after deliver. The aim of the submitted study was to evaluate LP-X results in children older than 6 months. 250 sera of 158 infants of all age groups were tested for presence of LP-X. The frequency of LP-X positive results was 85.7% in the group of infants younger than 24 weeks (91 samples of 45 infants were tested) whereas in the group of infants of an age between 24 weeks and 14 years (159 samples of 113 children were tested) 16.35% positive results were obtained. LP-X positive results showed the same clinical significance in the second group of children as it was demonstrated in adults elsewhere.